Doxorubicin (DOX) is a chemotherapeutic agent used for treatment of different cancers and its clinical usage is hindered by the oxidative injury-related cardiotoxicity. This work aims to declare if the harmful effects of DOX on heart can be alleviated with the use of Coenzyme Q10 (CoQ10) or L-carnitine. The study was performed on seventy two female Wistar albino rats divided into six groups, 12 animals each: Control group; DOX group (10mg/kg); CoQ10 group (200mg/kg); L-carnitine group (100mg/kg); DOX+CoQ10 group; DOX+L-carnitine group. CoQ10 and L-carnitine treatment orally started 5days before a single dose of 10mg/kg DOX that injected intraperitoneally (IP) then the treatment continued for 10days. At the end of the study, serum biochemical parameters of cardiac damage, oxidative stress indices, and histopathological changes were investigated. CoQ10 or L-carnitine showed a noticeable effects in improving cardiac functions evidenced reducing serum enzymes as serum interleukin-1 beta (IL-1 β), tumor necrosis factor alpha (TNF-α), leptin, lactate dehydrogenase (LDH), Cardiotrophin-1, Troponin-I and Troponin-T. Also, alleviate oxidative stress, decrease of cardiac Malondialdehyde (MDA), Nitric oxide (NO) and restoring cardiac reduced glutathione levels to normal levels. Both corrected the cardiac alterations histologically and ultrastructurally. With a visible improvements in α-SMA, vimentin and eNOS immunohistochemical markers. CoQ10 or L-carnitine supplementation improves the functional and structural integrity of the myocardium.
Keywords: Cardiotoxicity; CoQ10 and L-carnitine; Dox; Vimentin; eNOS.
Objective: To study the effects of resveratrol in neuronal structures in traumatic brain injury (TBI).
Study Design: Thirty rats were categorized as (1) control group (n=10), saline solution administered i.p. for 14 days, (2) TBI group (n=10), trauma induced by weight-drop model on brain, and (3) TBI+Resveratrol group (n=10), 15 minutes after injury the rats were given resveratrol (10 μmoL/kg/i.p.) for 14 days. At the end of the experiment the cerebellum was excised for routine paraffin tissue protocol. Blood samples were tested for serum biochemical markers (MDA, SOD, CAT, and GSH-x).
Results: SOD, GPx, and CAT values were lowest in the TBI group. MDA and histological scores of dilations in vessels, inflammation, degeneration in neurons, apoptosis in microglia, ADAMTS8, and GFAP expressions were highest in the TBI group. Sections of the control group showed normal cerebellar histology. The trauma group showed degenerated ganglion layer, pyknotic and apoptotic Purkinje cell nuclei. Vascular thrombus was seen in the substantia alba and substantia grisea. In the Trauma+Resveratrol group, most pa- thologies observed in the TBI group were improved. In the control group, GFAP protein was expressed in granular cells, axons, dendrites, Purkinje cells, and microglia cells. In the trauma group, increased GFAP expression was observed in glial processes, neurons, and Purkinje cells. In the Trauma+Resveratrol group, GFAP was expressed in molecular layer and glial processes. In the control group, ADAMTS-4 activity was observed in granulosa layer, glial cells, and Purkinje cells. In the trauma group, ADAMTS-4 expression was positive in Purkinje cells and glial cells. In the Trauma+ Resveratrol group, ADAMTS-4 was expressed in Purkinje cells, granular cells, and glial cells.
Conclusion: GFAP and ADAMTS-4 proteins may be involved in regeneration of damaged astroglial cells and other glial cells, Purkinje cells, and synaptic extensions. We suggest that antioxidative drugs such as resveratrol may be alternative target agents in neurological disease.
Keywords: ADAMTS-4, brain, cerebellum, GFAP, rat, resveratrol, traumatic brain injury
Liver ischemia/reperfusion injury, a setting in which the functional mass is ...Prof. Hesham N. Mustafa
Liver ischemia reperfusion is induced during sur-gical procedures like liver transplantation and re-section. Multiple mechanisms have been postulat-ed to liver damage following liver ischemia reperfu-sion injury, such as oxidative stress and inflamma-tory reactions. The present study declares the pos-sible mechanism of tadalafil, toward modulating the inflammatory response. Forty-eight rats were divided into 4 groups as follows; Sham group sub-jected to midline laparotomy only. Tadalafil group administered Tadalafil 10 mg/kg intraperitoneal 45 min before sham operation. I/R (Ischemia-reperfusion) group, rats undergo 60 min of hepatic ischemia followed by 60 min of reperfusion. Tada-lafil + I/R group rats undergo a similar pattern of I/R after the treatment with Tadalafil 10 mg/kg, 45 min before ischemia. At the end of the reperfusion, the blood samples were collected for estimation of biochemical markers including liver enzymes using colorimetric assay method and serum: TNF-α (tumor necrosis factor-α), IL-6 (interleukin 6) le-vels, ICAM- 1 (Intercellular Adhesion Molecule-1) were measured. Tissues were evaluated by semi-quantitative and morphometrical approaches. Ta-dalafil succeeded in restoring normal levels of liverenzymes and ameliorating the oxidative stress as evidenced by decreasing MDA and restoring redu-ced glutathione levels in liver tissue homogenate. Also, Tadalafil exhibits anti-inflammatory effects, as it significantly decreased the levels of TNF-α, IL6 and ICAM-1. The findings are supported by BCL-2, TNF-α immunomarkers. It is concluded that modulation of the inflammatory response might be one of the mechanisms of Tadalafil-mediated he-patoprotection, so it is recommended as an adju-vant therapy in liver surgery.Keywords: Ischemia/reperfusion injury – Oxidative stress – Apoptosis – TNF-α – BCL-2
Objective: To study the effects of resveratrol in neuronal structures in traumatic brain injury (TBI).
Study Design: Thirty rats were categorized as (1) control group (n=10), saline solution administered i.p. for 14 days, (2) TBI group (n=10), trauma induced by weight-drop model on brain, and (3) TBI+Resveratrol group (n=10), 15 minutes after injury the rats were given resveratrol (10 μmoL/kg/i.p.) for 14 days. At the end of the experiment the cerebellum was excised for routine paraffin tissue protocol. Blood samples were tested for serum biochemical markers (MDA, SOD, CAT, and GSH-x).
Results: SOD, GPx, and CAT values were lowest in the TBI group. MDA and histological scores of dilations in vessels, inflammation, degeneration in neurons, apoptosis in microglia, ADAMTS8, and GFAP expressions were highest in the TBI group. Sections of the control group showed normal cerebellar histology. The trauma group showed degenerated ganglion layer, pyknotic and apoptotic Purkinje cell nuclei. Vascular thrombus was seen in the substantia alba and substantia grisea. In the Trauma+Resveratrol group, most pa- thologies observed in the TBI group were improved. In the control group, GFAP protein was expressed in granular cells, axons, dendrites, Purkinje cells, and microglia cells. In the trauma group, increased GFAP expression was observed in glial processes, neurons, and Purkinje cells. In the Trauma+Resveratrol group, GFAP was expressed in molecular layer and glial processes. In the control group, ADAMTS-4 activity was observed in granulosa layer, glial cells, and Purkinje cells. In the trauma group, ADAMTS-4 expression was positive in Purkinje cells and glial cells. In the Trauma+ Resveratrol group, ADAMTS-4 was expressed in Purkinje cells, granular cells, and glial cells.
Conclusion: GFAP and ADAMTS-4 proteins may be involved in regeneration of damaged astroglial cells and other glial cells, Purkinje cells, and synaptic extensions. We suggest that antioxidative drugs such as resveratrol may be alternative target agents in neurological disease.
Keywords: ADAMTS-4, brain, cerebellum, GFAP, rat, resveratrol, traumatic brain injury
Liver ischemia/reperfusion injury, a setting in which the functional mass is ...Prof. Hesham N. Mustafa
Liver ischemia reperfusion is induced during sur-gical procedures like liver transplantation and re-section. Multiple mechanisms have been postulat-ed to liver damage following liver ischemia reperfu-sion injury, such as oxidative stress and inflamma-tory reactions. The present study declares the pos-sible mechanism of tadalafil, toward modulating the inflammatory response. Forty-eight rats were divided into 4 groups as follows; Sham group sub-jected to midline laparotomy only. Tadalafil group administered Tadalafil 10 mg/kg intraperitoneal 45 min before sham operation. I/R (Ischemia-reperfusion) group, rats undergo 60 min of hepatic ischemia followed by 60 min of reperfusion. Tada-lafil + I/R group rats undergo a similar pattern of I/R after the treatment with Tadalafil 10 mg/kg, 45 min before ischemia. At the end of the reperfusion, the blood samples were collected for estimation of biochemical markers including liver enzymes using colorimetric assay method and serum: TNF-α (tumor necrosis factor-α), IL-6 (interleukin 6) le-vels, ICAM- 1 (Intercellular Adhesion Molecule-1) were measured. Tissues were evaluated by semi-quantitative and morphometrical approaches. Ta-dalafil succeeded in restoring normal levels of liverenzymes and ameliorating the oxidative stress as evidenced by decreasing MDA and restoring redu-ced glutathione levels in liver tissue homogenate. Also, Tadalafil exhibits anti-inflammatory effects, as it significantly decreased the levels of TNF-α, IL6 and ICAM-1. The findings are supported by BCL-2, TNF-α immunomarkers. It is concluded that modulation of the inflammatory response might be one of the mechanisms of Tadalafil-mediated he-patoprotection, so it is recommended as an adju-vant therapy in liver surgery.Keywords: Ischemia/reperfusion injury – Oxidative stress – Apoptosis – TNF-α – BCL-2
Objective: A spinal cord injury (SCI) is damage to the spinal cord either from trauma, loss of its normal blood supply, or compression from tumor or infection. In this study we focused on alterations in the bladder tissue with angiogenic and apoptotic aspects after spinal cord injury.
Study Design: Twenty Wistar Albino rats were categorized as control and SCI groups. At T7-T9 vertebras, a steel rod was dropped from 10 cm to create a spinal cord injury under anesthesia. Rats were decapitated and spinal tissue was processed to measure malondialdehyde (MDA), glutathione (GSH), and myeloperoxidase (MPO).
Results: MDA, MPO, epithelial degeneration, vascular dilation, inflammation, VEGF, and APAF-1 expressions in the SCI group were statistically higher than those in the control group. GSH content of the SCI group was statistically lower than that in the control group. In the hematoxylin-eosin–stained sections of the control group, normal histology was observed in bladder tissue. In the SCI group, degeneration epithelial cells, thinned epithelium, increased fibrosis, dilated and congested blood vessels, and hyperplastic endothelial cells were observed. In the control group, VEGF expression was slightly observed in some epithelial cells and vascular cells. In the SCI group, VEGF expression was increased in inflammatory and vascular endothelial cells. For APAF-1 expression, the control group showed no expression. In the SCI group, APAF-1 expression was positive in degenerated epithelial cells and connective tissue cells.
Conclusion: It is thought that the urination reflex was affected due to increased inflammation in the bladder tissue, leading to alterations in the regulation and function of the muscles.
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
Does allicin combined with vitamin B-complex have superior potentials than al...Prof. Hesham N. Mustafa
BACKGROUND:
The current article aims to explore the protective potentials of α-tocopherol alone and the combination of allicin and vitamin B-complex against lead-acetate neurotoxicity on the cerebellar cortex.
MATERIALS AND METHODS:
Forty rats were divided into four groups (n=10). Group 1 was the control group. Group 2 received 10 mg/kg body weight (BW) of lead acetate. Group 3 was exposed to 10 mg/kg BW of lead acetate plus a combination of allicin (100 mg/kg BW) and vit. B-complex (40 mg/kg BW). Group 4 was administered lead acetate (10 mg/kg BW) and α-tocopherol (100 mg/kg BW). The animals received treatment for sixty days by oral gavage. All the groups were studied ultrastructurally and immunohistochemically with glial fibrillary acidic protein (GFAP).
RESULTS:
The affected groups revealed shrunken and degenerated Purkinje cells with irregular nuclei. The cytoplasm comprised several lysosomes, unhealthy mitochondria, and dilated Golgi saccules. The myelinated nerve fibers demonstrated breaking of the myelin sheaths, apparent vacuoles, and broad axonal spaces. Immunohistochemically, there was a tremendous surge in GFAP-positive astrocytes in the lead acetate-treated group. These histological and ultrastructural variations were ameliorated by the administration of α-tocopherol and the combination of allicin and vit. B complex. Moreover, an apparent decrease in the number of GFAP-positive astrocytes was obvious in the protected groups.
CONCLUSIONS:
Although both α-tocopherol and the combination of allicin and vit. B-complex can be used as possible adjuvant therapies to ameliorate nervous system ailments attributable to lead acetate, α-tocopherol showed more protective potential.
KEYWORDS:
Allicin; Astrocytes; GFAP; Myelin Figure; Oligodendrocyte; Purkinje cells
Effects of Different Proportions of Salvianolic Acid and Hydroxysafflor Yello...Premier Publishers
In this experiment, SD rats were injected intravenously with different proportions of test samples to observe the protective effect of intravenous injection of pituitrin (PIT) that induced myocardial ischemia in rats, and to determine the lipid peroxidation product of Malondialdehyde (MDA) in rat brain and the content and activity of Superoxide Dismutase (SOD), Creatine Kinase (CK) and Lactate Dehydrogenase(LDH). The role of the anti–myocardial ischemia model in the test and its optimal ratio were studied. Each group of test samples was injected through the tail vein and the Ⅱ lead ECG was traced. After injection for 30 min, the rats were sublingually injected with 2U/kg pituitrin, and Ⅱ lead ECG was recorded for 5 min. After 3 hours’ observation, the rats were sacrificed from the cervical spine, and the hearts were removed and placed in a refrigerator at –20 °C to measure various biochemical indicators. Intravenous injection of each group of tested products has the effect of reducing the content of MDA, LDH, and CK in myocardial tissue, which can enhance the activity of SOD in myocardial tissue, and has statistical significance compared with the model group (P≤0.05 or P≤0.01). Through comprehensive comparison, the A:B=1:10 dose group was found to have the best results.
Objective: A spinal cord injury (SCI) is damage to the spinal cord either from trauma, loss of its normal blood supply, or compression from tumor or infection. In this study we focused on alterations in the bladder tissue with angiogenic and apoptotic aspects after spinal cord injury.
Study Design: Twenty Wistar Albino rats were categorized as control and SCI groups. At T7-T9 vertebras, a steel rod was dropped from 10 cm to create a spinal cord injury under anesthesia. Rats were decapitated and spinal tissue was processed to measure malondialdehyde (MDA), glutathione (GSH), and myeloperoxidase (MPO).
Results: MDA, MPO, epithelial degeneration, vascular dilation, inflammation, VEGF, and APAF-1 expressions in the SCI group were statistically higher than those in the control group. GSH content of the SCI group was statistically lower than that in the control group. In the hematoxylin-eosin–stained sections of the control group, normal histology was observed in bladder tissue. In the SCI group, degeneration epithelial cells, thinned epithelium, increased fibrosis, dilated and congested blood vessels, and hyperplastic endothelial cells were observed. In the control group, VEGF expression was slightly observed in some epithelial cells and vascular cells. In the SCI group, VEGF expression was increased in inflammatory and vascular endothelial cells. For APAF-1 expression, the control group showed no expression. In the SCI group, APAF-1 expression was positive in degenerated epithelial cells and connective tissue cells.
Conclusion: It is thought that the urination reflex was affected due to increased inflammation in the bladder tissue, leading to alterations in the regulation and function of the muscles.
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
Does allicin combined with vitamin B-complex have superior potentials than al...Prof. Hesham N. Mustafa
BACKGROUND:
The current article aims to explore the protective potentials of α-tocopherol alone and the combination of allicin and vitamin B-complex against lead-acetate neurotoxicity on the cerebellar cortex.
MATERIALS AND METHODS:
Forty rats were divided into four groups (n=10). Group 1 was the control group. Group 2 received 10 mg/kg body weight (BW) of lead acetate. Group 3 was exposed to 10 mg/kg BW of lead acetate plus a combination of allicin (100 mg/kg BW) and vit. B-complex (40 mg/kg BW). Group 4 was administered lead acetate (10 mg/kg BW) and α-tocopherol (100 mg/kg BW). The animals received treatment for sixty days by oral gavage. All the groups were studied ultrastructurally and immunohistochemically with glial fibrillary acidic protein (GFAP).
RESULTS:
The affected groups revealed shrunken and degenerated Purkinje cells with irregular nuclei. The cytoplasm comprised several lysosomes, unhealthy mitochondria, and dilated Golgi saccules. The myelinated nerve fibers demonstrated breaking of the myelin sheaths, apparent vacuoles, and broad axonal spaces. Immunohistochemically, there was a tremendous surge in GFAP-positive astrocytes in the lead acetate-treated group. These histological and ultrastructural variations were ameliorated by the administration of α-tocopherol and the combination of allicin and vit. B complex. Moreover, an apparent decrease in the number of GFAP-positive astrocytes was obvious in the protected groups.
CONCLUSIONS:
Although both α-tocopherol and the combination of allicin and vit. B-complex can be used as possible adjuvant therapies to ameliorate nervous system ailments attributable to lead acetate, α-tocopherol showed more protective potential.
KEYWORDS:
Allicin; Astrocytes; GFAP; Myelin Figure; Oligodendrocyte; Purkinje cells
Effects of Different Proportions of Salvianolic Acid and Hydroxysafflor Yello...Premier Publishers
In this experiment, SD rats were injected intravenously with different proportions of test samples to observe the protective effect of intravenous injection of pituitrin (PIT) that induced myocardial ischemia in rats, and to determine the lipid peroxidation product of Malondialdehyde (MDA) in rat brain and the content and activity of Superoxide Dismutase (SOD), Creatine Kinase (CK) and Lactate Dehydrogenase(LDH). The role of the anti–myocardial ischemia model in the test and its optimal ratio were studied. Each group of test samples was injected through the tail vein and the Ⅱ lead ECG was traced. After injection for 30 min, the rats were sublingually injected with 2U/kg pituitrin, and Ⅱ lead ECG was recorded for 5 min. After 3 hours’ observation, the rats were sacrificed from the cervical spine, and the hearts were removed and placed in a refrigerator at –20 °C to measure various biochemical indicators. Intravenous injection of each group of tested products has the effect of reducing the content of MDA, LDH, and CK in myocardial tissue, which can enhance the activity of SOD in myocardial tissue, and has statistical significance compared with the model group (P≤0.05 or P≤0.01). Through comprehensive comparison, the A:B=1:10 dose group was found to have the best results.
Antioxidant potentials of tannic acid on lipid peroxidation induced by severa...Premier Publishers
Various prospective studies have indicated the antioxidant potency of tannic acid in several models. However, there is no clear-cut evidence revealing that the reported antioxidant properties of tannic acid remains potent regardless of the lipid sources and pro-oxidants employed for the oxidative assault. Hence, this study sought to investigate the antioxidant properties of tannic acid against cerebral and hepatic lipid peroxidation induced by several pro-oxidants (Iron (II) sulfate, Sodium nitroprusside, cyclophosphamide and acetaminophen) in vitro. Rats were decapitated under mild ether anesthesia and the tissues were rapidly dissected, placed on ice, weighed and immediately homogenized in cold 50 mM Tris-HCl, pH 7.4 (1/10, w/v). The homogenates were centrifuged for 10 min at 4000 g to yield a pellet that was discarded and a low-speed supernatant (S1). Our results indicated that Fe (II) showed the highest pro-oxidative effects in both tissues lipids. Furthermore, tannic acid demonstrated potent inhibitory effects against lipid peroxidation in both tissues lipids regardless of the pro-oxidant employed. To this end, there is a dire need to exploit the protective benefits of tannic acid as a potential exogenous antioxidant against lipid peroxidation with a view to providing solution to the global oxidative stress menace.
A Study on the Toxic Effect of Different Doses of Diclofenac Sodium on the De...Prof. Hesham N. Mustafa
SUMMARY: The toxic effects of different doses of diclofenac sodium (DS) on the kidney on the postnatal period (0-7 days) by
morphometrical and immunohistochemical methods were investigated. For this purpose, 15 female adult wistar albino rats were used and
divided into 5 main groups. Group Ia served as normal control, physiologic group Ib received normal saline, group II received low dose (3.9
mg/kg), group III received medium dose (9 mg/kg) and group IV received high dose (18 mg/kg). Male offspring’s from 0-7 days after birth
were used in this study. On the 8th day of postnatal life, all animals were anesthetized. Then, the kidney samples were analyzed. Haematoxylin
and eosin staining showed degeneration and necrosis, apparent atrophy of the glomeruli, mononuclear cell infiltration, congested vessels,
increased fibrous tissue and distortion of the proximal convoluted tubules with interruption of the brush margin of the DS treated group.
Increased level of Caspase-3 and upregulation of TNF-α with different doses of DS. In light of our findings, DS may lead to adverse effects
that are dose-dependent in the prenatal subjected kidney to this drug.
KEY WORDS: Diclofenac sodium; Proximal convoluted tubules; Apoptosis;Cyclooxygenase.
A study on the toxic effect of different doses of Diclofenac sodium on the de...Prof. Hesham N. Mustafa
The toxic effects of different doses of diclofenac sodium (DS) on the kidney on the postnatal period (0-7 days) by morphometrical and immunohistochemical methods were investigated. For this purpose, 15 female adult wistar albino rats were used and divided into 5 main groups. Group Ia served as normal control, physiologic group Ib received normal saline, group II received low dose (3.9 mg/kg), group III received medium dose (9 mg/kg) and group IV received high dose (18 mg/kg). Male offspring’s from 0-7 days after birth were used in this study. On the 8th day of postnatal life, all animals were anesthetized. Then, the kidney samples were analyzed. Haematoxylin and eosin staining showed degeneration and necrosis, apparent atrophy of the glomeruli, mononuclear cell infiltration, congested vessels, increased fibrous tissue and distortion of the proximal convoluted tubules with interruption of the brush margin of the DS treated group. Increased level of Caspase-3 and upregulation of TNF-α with different doses of DS. In light of our findings, DS may lead to adverse effects that are dose-dependent in the prenatal subjected kidney to this drug.
Keywords : Diclofenac sodium; Proximal convoluted tubules; Apoptosis; Cyclooxygenase.
Background and objectives: the 5’nucleotidase (5’NT) is one of
hydrolytic enzymes present in different organs which catalyze
hydrolysis of 5’ nucleotides to ribonucleosides and phosphate.
Malondialdehyde (MDA) is the end product of lipid peroxidation
by oxidative stress (free radicals).
The aim of present study was to measure the serum activity of
5’NT, and MDA concentration in breast tumors.
Material and method: A prospective study was carried out from
May to December 2013 by clinical biochemistry department in
College of Medicine-Hawler Medical University on (30) healthy
female individuals, (group 1) and (30) females with breast tumor
(group 2).
Results: The mean value of serum MDA was significantly higher
in females with breast tumors (group 2), than that of healthy
female individuals, (group 1) (p<0.01),><0.01).
Conclusion: Based on findings of the present study it can be
concluded that breast tumors can cause release of the enzyme
5’NT from tumor cells, and lipid peroxidation by reactive oxygen
species (ROS), which cause elevation of MDA.
Ameliorative potential of the quercetin on lead-induced testicular damage mor...Prof. Hesham N. Mustafa
Background
Quercetin, a naturally occurring flavonoid known for its potent antioxidant properties, has been investigated for its potential in counteracting the harmful effects of lead (Pb) toxicity, which induces apoptosis and oxidative damage in various human tissues. This study aims to assess the reparative effects of quercetin on lead-induced testicular damage.
Methods
Four groups, each comprising ten adult male albino rats, were randomly assigned as follows: Quercetin group, Pb group, Pb + Quercetin group, and control group. All treatments were administered orally via gavage daily for a duration of 30 days. Evaluation of sex hormone levels (serum testosterone, FSH, and LH), cytokines and inflammatory mediators (IL-1β, TNF-α, MCP-1), lead concentration, oxidative and antioxidant stress markers (superoxide anion [O2−], MDA, SOD, CAT, GSH), and sperm characteristics were carried out.
Results
The results demonstrated a significant decline in sex hormones and antioxidants, accompanied by an increase in lead concentrations, cytokines, inflammatory mediators, and oxidative stress indicators (O2−, MDA), while SOD, CAT, and GSH levels were reduced. The Pb-intoxicated group exhibited a substantial increase in dead and abnormal sperm, along with significant reductions in sperm concentration and motility. Morphometrically, a marked decrease was observed in spermatogonia, primary spermatocytes, spermatids, and sertoli cells per seminiferous tubule, as well as epithelial height. Furthermore, coadministration of quercetin exhibited notable benefits. It significantly elevated testosterone levels (P < 0.001), testicular SOD, CAT, and GSH activities, while decreasing MDA levels (P < 0.001). Quercetin also mitigated the deleterious effects of lead toxicity on sperm parameters and restored morphometric variations, including epithelial height.
Conclusions
Quercetin supplementation alongside lead exposure showed a potential for ameliorating degenerative changes caused by lead toxicity in the testicles. This cotreatment effectively reduced oxidative stress, cytokine levels, inflammatory mediators, and restored biochemical alterations, thereby improving morphometric parameters.
The pattern of branching and intercommunications of the musculocutaneous nerv...Prof. Hesham N. Mustafa
Background:
The aim of the present work was to provide evidence about the anatomical variations as regard the origin, distribution, and branching pattern of the musculocutaneous nerve (MCN).
Materials and methods:
Brachial plexus was dissected in 40 upper limbs of 20 male adult cadavers. The pattern of the musculocutaneous nerve was photographed by a digital camera.
Results:
The location and length of the nerve branches between left and right arms were recorded and statistically analyzed. In (90%) of specimens the MCN originates from the lateral cord of the brachial plexus, in (5%) it arose from the median nerve (MN), while in the remaining (5%) specimen, it was absent. The musculocutaneous nerve pierced the coracobrachialis muscle in 90% of specimens, and in the remaining (10%) did not pierce it. The motor branches to biceps brachii muscle were categorized into: Type 1 (90%): one branch that divides to supply the two heads of biceps; Type 2 (5%): double branches, innervating each head of biceps separately. The motor branches to brachialis muscle were categorized into: Type 1 (82.9%): one branch; Type 2 (14.2%): double branches and Type 3 (2.9%): three branches that innervating brachialis muscle. Communications between the MCN and the MN were observed in 35% of specimens.
Conclusions:
The knowledge of the common and uncommon musculocutaneous nerve variations is important especially to the surgeons for carrying out surgical procedures in axilla and arm.
Morphohistometric analysis of the effects of Coriandrum sativum on cortical a...Prof. Hesham N. Mustafa
Objective: Natural compounds can act as metal chelators and oxygen free radical scavengers, which allows them to be used as bioactive antagonists to heavy metals neurotoxicity. The aim of the study to analyze the morphometric effects of Coriandrum sativum (C. sativum) on lead-induced neurotoxicity.
Materials and Methods: Forty Sprague-Dawley albino rats were divided into four equal groups (ten in each group): control group; coriander group: received aqueous C. sativum extracts (600 mg/kg BW for 60 days orally); lead (Pb) group: received a daily dose of lead acetate (Pb) (10 mg/kg BW for 60 days orally); Pb+ coriandrum group: received: aqueous C. sativum extract (600 mg/kg BW) prior to 10 mg/kg BW of Pb. The following parameters malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were measured. Layers thickness and nuclei density were analyzed.
Results: Lead levels in blood and tissues were decreased significantly in the Pb group and those findings were corrected significantly (p=0.001) with C. sativum addition. Data exhibited an increase in oxidative stress marker MDA and a decrease in antioxidant enzymes activities (SOD, CAT, and GPx) significantly in the Pb group and those effects were reversed significantly (p=0.001) by C. sativum administration. The cerebellar cortex and all layers of the somatosensory cortex thickness and nuclei density were diminished significantly in the Pb group. The morphometrical measurements were corrected significantly (p=0.001) by C. sativum.
Conclusion: From the findings of the current study, Pb caused noticeable structural and functional variations in the cerebellar cortex and somatosensory cortex. C. sativum corrected these parameters as it possesses chelating and antioxidant potentials.
Background:
The anterolateral ligament (ALL) is a true well-defined ligament in the knee first described in 1879 by Segond. After the work of Claes et al., several studies were conducted about biomechanics and its role in stability of the knee. The anatomical existence of the ALL has been studied by and various radiographic diagnostic modalities and in cadavers. It originates from lateral femoral epicondyle and is inserted between Gerdy’s tubercle and the fibular head. There has been controversy about the existence of ALL in pediatric patients. The aim of this work was to confirm the presence of ALL in pediatric patients by using MRI.
Materials and Methods:
We reviewed the knee MRI scans of 100 pediatric patients (ages between one and 12 yr) who had no knee injury or congenital deformity and had been evaluated by an expert radiologist.
Results:
The ALL was detected in 90% of the pediatric patients with the use of MRI.
Conclusions:
The main finding of this study was that ALL can be seen in pediatric patients using MRI. Despite numerous studies, additional research is needed to further define the role of the ALL in knee function.
Level of Evidence:
Level IV.
Protective effect of garlic extract against maternal and fetal cerebellar dam...Prof. Hesham N. Mustafa
Background: In spite of its industrial usefulness and varied daily uses, lead (Pb) pollution is a widespread ecological problem that faces the humans in the 21th century. Pb was found to produces a wide range of toxic effects including neurotoxicity especially to the developing and young offspring. Recently, the utilisation of herbal plants has received a significant attention where there has been rising awareness in their therapeutic use; among these is the garlic. In light of the above, the current study is designed experimentally in female pregnant rats in order to investigate the beneficial role of garlic extract in the protection from the maternal and foetal cerebellar damage produced by administration of different doses of Pb during pregnancy.
Materials and methods: Positively pregnant female rats were divided into five groups; one control group, two Pb-treated groups (exposed to 160 and 320 mg/kg b.w. of Pb, respectively) and two groups treated with both Pb and garlic (exposed to Pb as previous groups together with 250 mg/kg b.w./day of garlic extract). Treatments started from day 1 to day 20 of pregnancy, where the mother rats of different experimental groups were sacrificed to obtain the foetuses. Pb level in the maternal and foetal blood and cerebellum was estimated by spectrophotometry. Specimens of the cerebellum of different mother and foetal groups were processed to histological and immunohistochemical staining for microscopic examination.
Results: The results showed that administration of Pb to pregnant rats resulted in a dose-dependent toxicity for both mothers and foetuses in the form of decrease in maternal weight gain, placental and foetal weights, brain weight and diminished foetal growth parameters, which were prominent in rat's group treated with larger dose of Pb. In Pb-treated rats, Pb level in blood and cerebellum was high when compared with the control group. The histopathological examination of the cerebellum of treated dams and foetuses showed marked alterations mainly in the form of Purkinje cell degeneration and lack of development of foetal cerebellum. Co-treatment of garlic extract along with Pb resulted in a significant decrease in Pb levels as compared with those treated with Pb alone with improvement of the histopathological changes.
Conclusions: This study was useful in evaluating the hazardous effects of uncontrolled use of Pb in general and in assessing the developmental and neurotoxicity of foetuses due to exposure during pregnancy in particular. Co-administration of garlic has beneficial effects in amelioration of Pb-induced neurotoxicity and reversing the histopathological changes of the cerebellum of mother rats and foetuses. (Folia Morphol 2018; 77, 1: 1-15).
Keywords: Purkinje cells; garlic; glial fibrillary acidic protein; lead.
One year mortality rate after hip fracture in the western region of saudi ara...Prof. Hesham N. Mustafa
Background:
The mortality rate of elderly patients who sustain a hip fracture is high compared to the general population. Identifying risk factors can help predict patients at risk of hip fracture to reduce the mortality rate. No studies have shown the mortality rate of patients with hip fractures in the western region of Saudi Arabia. Therefore, this study aimed to identify the risk factors associated with the mortality of patients with hip fractures admitted to the King Abdulaziz Hospital and compare the results with other studies.
Methods:
The mortality rate (within 1 yr or less) in 177 patients over the age of 60 yr who were admitted to the university hospital between July, 2007, and September, 2012, with hip fractures was retrospectively studied. The patients were assessed with regard to gender, age, type of hip fracture, and type of surgical intervention.
Results:
The overall mortality rate 1 yr after hip fracture was 12.43%, and the mean age was 77.77 yr old. The risk factors most associated with mortality were as follows: advanced age (71 to 80 and 81 to 90 yr old), male, peritrochanteric (extracapsular) fracture, and operative fixation with dynamic hip screw.
Conclusions:
The mortality rate of patients with hip fractures within 1 yr has a high-risk potential, especially for male patients over 71 yr of age with peritrochanteric (extracapsular) fractures. Surgical treatment with dynamic hip screw also was shown to be a risk factor between the different treatment options.
Level of Evidence:
Level IV.
Biomarkers of Systemic Lupus Erythematosus and Systemic Sclerosis diseases ac...Prof. Hesham N. Mustafa
Systemic Lupus Erythematosus (SLE) and systemic sclerosis (SSc) are systemic inflammatory autoimmune disorders characterized by a large spectrum of clinical and laboratory features. The aim of the present study was to investigate the possible use of serum level of soluble intercellular adhesion molecule-1(sICAM-1) and soluble interleukin-2 receptor (sIL-2Ra) as biomarkers for monitoring of SLE and SSc disease activity. Moreover, it aimed to compare the specificity and sensitivity as well as cut-off value of both biomarkers in a sample of Egyptian patients. 50 SLE patients, 30 SSc patients and 60 age and sex matched healthy controls were enrolled in our study. sICAM-1and sIL-2Ra were measured in serum samples obtained from all participants. In addition to Erythosedimentation rate (ESR), complete blood count (CBC), Antineuclearantibodies (ANA) estimation, disease activity of both diseases were also assessed. sICAM-1and sIL-2Ra levels were higher in SLE and SSc patients versus control. Both parameters are correlated with each other as well as the activity parameters. A cut-off levels of 455.59 (ng/ml) &2525935 (pg/ml) in both SLE & SSs respectively was observed with the highest specificity and sensitivity. It could be concluded that sICAM-1 and sIL-2Ra are noninvasive biomarkers for SLE and SSc that could play a pathophysiologic role in development and progression of both diseases. Moreover, sICAM-1 and sIL-2Ra are correlated with the disease activity at cut-off values of 455.59 (ng/ml) & 2525935(pg/ml) respectively.
Morphohistometric study of the ligamentum flavum in cervical,thoracic and lum...Prof. Hesham N. Mustafa
ABSTRACT Anatomic characterization and fine structure of the human ligamentum flavum (LF), especially at different spinal levels, represent an attractive focus for the scientific and surgical application. Descrip-tive anatomical and structural study of LF at the cervical, thoracic and lumbar levels of the vertebral column in human cadavers is carried out here. The aim of the work is to clarify the anatomical features and fine structural differences in the human LF at different vertebral levels (cervical, thoracic and lumbar). Specimens of vertebral column were ob-tained from 34 human preserved cadavers. Their average age ranged between 56 and 69 years. Morphometric parameters including height, width and thickness of the ligament flavum at the mid-levels of cervical, thoracic and lumbar regions were measured. Sections obtained from different levels were stained with different stains. Morpho-metric measurements involved the relative elastic area, relative collagen area, elastic area% and collagen area% were measured.The results of the height, width and thickness of the LF at different spinal levels showed gradual increase in their mean values respectively. The LF midline gaps were found in the cervical, thoracic and lumbar regions. The morphometrical measure-ments showed that the average elastic area was highest in the cervical region and lowest in the tho-racic region. In the lumbar region, the percentages of both elastic area and the collagen area were nearly the same. The characterization of morpho-logical and histological aspects of the LF at differ-ent spinal levels will be of great importance for ap-plications in spinal surgery, biomechanical and physical rehabilitation of vertebral column.Keywords: Ligamentum Flavum – Spinal – Collagen and elastic fibers
Correlation between acl injury and involvement of the anterolateral ligament ...Prof. Hesham N. Mustafa
Background:
Clinical testing has demonstrated the role of the anterolateral ligament (ALL) in controlling anterolateral laxity and knee instability at high angles of flexion. Few studies have discussed the association between an anterior cruciate ligament (ACL) injury and ALL injury, specifically after residual internal rotation and a post-ACL reconstruction positive pivot-shift that could be attributed to ALL injury. The goal of this study was to assess the correlation between ALL injury and ALL injury with concomitant ACL injury using MRI.
Material and Methods:
This was a retrospective study of 246 patients with unilateral ACL knee injuries from a database that was reexamined to identify whether ALL injuries occurred in association with ACL injuries. We excluded the postoperative reconstructed cases. The charts were reviewed on the basis of the presence or absence of diagnosed ACL injury with no regard for age or sex.
Results:
Of the 246 patients with ACL injury, there were 165 (67.1%) patients with complete tears, 55 (22.4%) with partial tears, and 26 (10.6%) with sprains. There were 176 (71.5%) patients with ALL and associated ACL injuries, whereas 70 (28.5%) did not have associated ACL injuries. There was a significant statistical relationship between ACL and ALL injuries (P<0.0001).
Conclusions:
There is high incidence of ALL tears associated with ACL injuries. Clinicians should be aware of this injury and consider the possibility of simultaneous ALL and ACL repair to prevent further knee instability.
Level of Evidence:
Level IV.
Neuro-amelioration of cinnamaldehyde in aluminum-induced Alzheimer’s disease ...Prof. Hesham N. Mustafa
Aluminum (Al) is a neurotoxic substance which has played an important role in the etiology, pathogenesis, and development of amyloid-β (Aβ) plaques. This study was carried out to evaluate the neuroprotective effect of aqueous cinnamon extract against aluminum chloride (AlCl3)-induced Alzheimer’s disease. Forty adult male albino rats, randomly divided into four equal groups. Control group; ACE200 group administered aqueous cinnamon extract (ACE) orally; AlCl3 group received daily intraperitoneal (i.p.) injection of AlCl3 for 60 days to induce neurotoxicity and AlCl3 + ACE200 group received a combination of AlCl3 and ACE in the same dose and route as previous groups. Aluminum administration significantly enhanced the memory impairment and the Aβ formation in the rat model. The cerebellum exhibited a significant reduced number of Purkinje cells, marked decrease in the density of dendritic arborization and prominent perineuronal spaces in the molecular layer. There was loss of dendritic spines, neurofibrillary degeneration, and appearance of neuritic plaques. Concomitant administration of AlCl3 and ACE displayed an observable protection against these changes with progressive improvement in memory and intellectual performance. In conclusion, ACE may play a protective role against formation of amyloid-β plaques in cerebellum.
Analytical Study of Clinicopathological Data of Saudi Patients with Osteoarth...Prof. Hesham N. Mustafa
SUMMARY: Knee osteoarthritis (OA) is a common disabling disease. Epidemiological studies have revealed various risk
factors for OA, including sex, aging, obesity, occupational illnesses, and chronic diseases. Here we evaluate the clinical, pathological,
and radiological findings of knee OA in a subset of Saudi patients who were subjected to total knee replacement (TKA). The study
population included 30 Saudi patients with knee OA who were operated by TKA (from June 2014 to December 2015) in the Department
of Orthopedics, Faculty of Medicine, King Abdulaziz University, Saudi Arabia. Patient’s clinical and radiological data were collected
from the hospital files. Pathological examination of the excised superior articular surface of tibia and femoral condyles were done.
Pearson Chi-squared analysis was used to test for differences between the variables in associated risk factors. There were more women
than men. Sixty per cent of patients were older than 60 years [mean age, 59.2 (females) and 61.7 (men) years-old]. All patients exceeded
obesity class 1, with females being more obese than males. Pathological examination of the superior articular surface of tibia and femoral
condyles showed high score lesions, which was more apparent in females than in males. Radiological findings showed that most lesions
were high grade. The findings of this study will help to understand the pathogenesis of OA and improve treatment decision making
relevant to TKA in knee OA in Saudi Arabia and elsewhere.
KEY WORDS: Osteoarthritis; Knee; Arthroplasty.
Thymoquinone ameliorates oxidative damage and histopathological changes of de...Prof. Hesham N. Mustafa
ABSTRACT
Lead (Pb) toxicity is known to be a chief environmental health issue, especially for pregnant
women and young children. Today, the use of medicinal herbs in the treatment of many diseases
and different toxic agents has become highly accepted due to their effectiveness and lower costs.
Thymoquinone (TQ), which is extracted from Nigella sativa seeds, is a potent antioxidant and anti
inflammatory agent. This study was designed to explore the optional protectivity of TQ against
maternal and fetal oxidative stress and brain damage induced by Pb administration. Pregnant
rats were distributed into seven groups: control group, TQ group, DMSO group, two groups Pb
treated (160 and 320 ppm), and two groups Pb-treated (160 and 320 ppm) co-treated with TQ.
Administration started from gestation day 1 (GD1) to day 20 (GD20) through oral gavage once
daily. Lead administration caused a dose-dependent toxicity for both mothers and fetuses. Also,
the histopathological assessment of the brains from Pb-treated groups showed marked altera
tions. Co-treatment of with TQ and Pb caused a significant decrease in Pb levels as compared
with those treated with Pb alone and amelioration of histopathological changes in the brains. It
was concluded that co-treatment of TQ along with gestational Pb exposure could mitigate the
effects against Pb-induced maternal and fetal neurotoxicity.
KEYWORDS
Lead; oxidative stress; brain;
Thymoquinone; fetal toxicity
Zingiber Officinale Alleviates Maternal and Fetal Hepatorenal Toxicity Induce...Prof. Hesham N. Mustafa
This study was designed to address the protective effects of Zingiber officinale on the toxic outcomes of prenatal Cadmium administration on pregnancy outcome. Pregnant female Sprague-Dawley rats were randomly divided into four groups (eight rats/each), control group received distilled water, 2nd group treated with 8.8 mg of CdCl2/kg b. wt, 3rd group treated with 250 mg of Zingiber officinale/kg b. wt, and 4th group treated with 250 mg of Zingiber officinale/kg b. wt, followed by 8.8 mg of CdCl2/kg b.wt. Daily body weight of pregnant was recorded from GD1-GD20, and then pregnant rats were sacrificed at GD20. Samples of maternal and fetal livers and kidneys were processed for histological examination. Administration of Cd to pregnant rats showed adverse effects on pregnant mothers and their fetuses; reduced maternal weight gain, reduced absolute organ weights, reduced fetal growth parameters and placental weights together with altered histological appearance of the maternal and fetal livers and kidneys. While co-administration of Zingiber officinale showed an improvement of these toxic alterations. Zingiber officinale through its antioxidant activity could be beneficial against toxic outcomes of Cd exposure during pregnancy.
Evaluation of the safety of conventional lighting replacement by artificial d...Prof. Hesham N. Mustafa
Background
Short morning exposure to high illuminance visible electromagnetic radiations termed as artificial daylight is beneficial for the mental health of people living in geographical areas with important seasonal changes in daylight illuminance. However, the commercial success of high illuminance light sources has raised the question of the safety of long hour exposure.
Methods
We have investigated the effect of the replacement of natural daylight by artificial daylight in Swiss mice raised under natural lighting conditions. Mice were monitored for neurotoxicity and general health changes. They were submitted to a battery of conventional tests for mood, motor and cognitive functions’ assessment on exposure day (ED) 14 and ED20. Following sacrifice on ED21 due to marked signs of neurotoxicity, the expression of markers of inflammation and apoptosis was assessed in the entorhinal cortex and neurons were estimated in the hippocampal formation.
Results
Signs of severe cognitive and motor impairments, mood disorders, and hepatotoxicity were observed in animals exposed to artificial daylight on ED20, unlike on ED14 and unlike groups exposed to natural daylight or conventional lighting. Activated microglia and astrocytes were observed in the entorhinal cortex, as well as dead and dying neurons. Neuronal counts revealed massive neuronal loss in the hippocampal formation.
Conclusions
These results suggest that long hour exposure to high illuminance visible electromagnetic radiations induced severe alterations in brain function and general health in mice partly mediated by damages to the neocortex-entorhinal cortex-hippocampus axis. These findings raise caution over long hour use of high illuminance artificial light.
The Ameliorative Potential of Dexmedetomidine and Benincasa Cerifera Extract ...Prof. Hesham N. Mustafa
Renal ischemia/reperfusion injury (IRI) represents the main reason for acute kidney injury (AKI). Dexmedetomidine (Dex) and Benincasa cerifera (BC) have wide benefits due to their anti-inflammatory and antioxidant properties. This study aims to illustrate the protective effects of BC and Dex on renal IRI in a diabetic model. Sixty adult male albino rats (Wistar strain), weighing 250–300 g, were included in the study. The rats were divided into four groups, as follows: sham group: (non-diabetic); diabetes mellitus (DM) + IRI group: streptozotocin (STZ)-induced diabetic rats exposed to renal IRI on day 30 after diagnosis of diabetes; DM + IRI + BC group: STZ-induced diabetic rats treated with BC (500 mg/kg) for 30 days after diagnosis of diabetes, then exposed to renal IRI; and DM + IRI + Dex group: STZ-induced diabetic rats treated with Dex (100 µg/kg intraperitoneally) 5 min before induction of ischemia on day 30 after diagnosis of diabetes, then exposed to renal IRI. Biochemical parameters, histopathological examination, and immunohistochemical markers were evaluated. A significant improvement in the biochemical, histopathological, and immunohistochemical parameters were observed in the DM + IRI + BC group, while the DM + IRI + Dex group showed improvements in renal IRI and dyslipidemia. The present study demonstrated that oxidative stress plays a chief role in renal IRI in the STZ-induced diabetic model. Treatment with BC achieved excellent ameliorative effects, while treatment with DEX improved renal IRI.
Keywords:
Diabetes; Dexmedetomidine; Ischemia/Reperfusion; Oxidative Stress
Beneficial Effects of Curcumin Inmaternal and Fetal Oxidativestress and Brain...Prof. Hesham N. Mustafa
This study was planned to explore the protective role of curcumin (Cur) against maternal and fetal oxidative stress and cerebral damage induced by lead (Pb) during pregnancy. Positively pregnant female rats were divided into seven groups: control group, Cur group (300 mg/kg of Cur/b.wt.), DMSO group (50% DMSO), two Pb-treated groups (exposed to 160 and 320 mg/kg b.wt./day of Pb acetate, respectively), and two groups treated with both Pb and Cur (exposed to Pb as previous groups together with 300 mg/kg b.wt./day of Cur). Treatments through oral gavage once a day started from gestation day 1 (GD1) till day 20 (GD20), where the mother rats of different experimental groups were sacrificed to obtain the fetuses. Different chemical parameters were assessed. Brain specimens of mother and fetal groups were processed with examination. The results displayed that Pb administration to pregnant rats resulted in a dose-dependent toxicity for both mothers and fetuses. Also, there was a significant rise in lipid peroxidation and decreased antioxidant enzyme activities in the brains of the different Pb-treated groups. The histological examination of the brain of treated dams and fetuses showed marked alterations. Co-treatment of Cur along with Pb caused a significant decrease in Pb levels as compared with those treated with Pb alone, improving the oxidative condition with amelioration of the brain’s histopathological changes. Co-administration of Cur could have ameliorative effect against Pb-induced neurotoxicity through the reduction of oxidative stress and reversal of histopathological changes.
Keywords:
Lead; Oxidative Stress; Brain; curcumin; Fetal toxicity
Immunohistochemical Study of the Ameliorative Effect of Vitamin E on Liver Re...Prof. Hesham N. Mustafa
The liver is almost unique in its capacity for regeneration after hepatectomy but the exact mechanisms are not yet fully clarified. Antioxidants have been shown to promote liver regeneration after major hepatectomy. The present study evaluated the ameliorative effect of vitamin E administration on the liver regeneration after different periods of partial hepatectomy (PH) in rats. Fifty-six adult male albino rats were divided into three groups: Control sham operated group; partially hepatectomized group which were divided into three subgroups sacrificed at 1day, 3 days and 7days after the operation respectively; Partially Hepatectomized group with vitamin E pretreatment before PH where the rats were given a daily oral dose of vitamin E until the time of sacrifice of the rats. Immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and labeling index were demonstrated. After PH, the PCNA positive hepatocytes and the PCNA labeling indices were significantly high after the 1st day and then much decreased after the 3rd day, to be followed by a slight increase at the 7th day. Vitamin E pretreatment in PH rats resulted in a decrease in PCNA positive cells and its labeling indices in the 1st day with a gradual increase in the 3rd and 7th days. Vitamin E has an inhibitory effect in the first 24 hours on liver regeneration followed by stimulatory effect at the third and seventh days after PH. These data indicated that vitamin E pretreatment has an important role in regulation and enhancement of liver regeneration after PH.
Keywords:
Immunohistochemistry; Labeling Index; Vitamin E; Partial Hepatectomy; PCNA
The role of curcumin in streptozotocin induced hepatic damage and the trans-d...Prof. Hesham N. Mustafa
Diabetic patients frequently suffer from non-alcoholic steatohepatitis. The current study aimed to investigate the role of curcumin and the response of hepatic stellate cells in streptozotocin (STZ)-induced hepatic damage. Sixty male rats were divided into three groups. The normal control injected with a citrate buffer vehicle and the diabetic control group which was injected intraperitoneally (IP) with a single-dose of streptozotocin (50mg/kg body weight) and a diabetic group was treated with an oral dose of curcumin at 80 mg/kg body weight daily for 60 days. Curcumin effectively counteracts oxidative stress-mediated hepatic damage and improves biochemical parameters. Alpha-smooth muscle actin (α-SMA) was significantly reduced, and insulin antibodies showed strong positive immunoreactivity with curcumin administration. These results optimistically demonstrate the potential use of curcumin, which is attributed to its antiradical/antioxidant activities and its potential β-cell regenerative properties. Also, it has the capability to encourage the trans-differentiation of hepatic stellate cells into insulin-producing cells for a period of time. In addition, as it is an anti-fibrotic mediator that inhibits hepatic stellate cell activation and the transition to myofibroblast-like cells, this suggests the possibility of considering curcumin's novel therapeutic effects in reducing hepatic dysfunction in diabetic patients.
Potential Alleviation of Chlorella vulgaris and Zingiber officinale on Lead-I...Prof. Hesham N. Mustafa
Natural products were studied to combat reproductive alterations of lead. The current work
aimed to disclose the efficacy of Chlorella vulgaris and Zingiber officinale to alleviate lead
acetate induced toxicity. Sixty adult male Wistar rats were distributed into four groups.
Group 1 was considered control, group 2 received 200 mg/l PbAc water, group 3 received 50
mg/kg/rat of C. vulgaris extract and 200 mg/l PbAc water, and group 4 received 100
mg/kg/rat of Z. officinale and 200 mg/l PbAc water for 90 days. Testis samples were subjected
to ultrastructural examination. It was observed that PbAc caused degenerative alterations in
the spermatogenic series in many tubules, with a loss of germ cells and vacuoles inside the
cytoplasm and between the germ cells. Mitochondria exhibited ballooning, with lost cristae
and widening of the interstitial tissue, while nuclear envelopes of primary spermatocytes
were broken up, and axonemes of the mid-pieces of the sperms were distorted. With the
treatment with C. vulgaris or Z. officinale, there were noticeable improvements in these
modifications. It was concluded that both C. vulgaris and Z. officinale represent convincing
medicinal components that may be used to ameliorate testicular toxicity in those exposed to
lead in daily life with superior potentials revealed by C. vulgaris due to its chelating action.
Key words: Chlorella vulgaris, lead acetate, ultrastructure, Zingiber officinale.
The ameliorative potential of hyphaene thebaica on streptozotocin induced dia...Prof. Hesham N. Mustafa
Background: Diabetic nephropathy (DN) is the leading cause of end-stage renal
disease. The aim of the current study is to investigate the possible beneficial effects
of Hyphaene thebaica in DN.
Materials and methods: For this, 50 male albino rats were divided into five
groups: group I — represented the control group; group II — received Hyphaene thebaica extracts of 150 mg/kg BW by oral gavage for 6 weeks; group III
— received single intraperitoneal injections of streptozotocin (50 mg/kg BW)
to induce type-2 diabetes mellitus; group IV (protective) — diabetic rats received Hyphaene thebaica extract (150 mg/kg BW) orally for 6 weeks; group V
(curative) — received Hyphaene thebaica extract (150 mg/kg BW) orally after the
diagnosis of DN.
Results: In the DN protected group, blood glucose, urea, and creatinine decreased
significantly, while insulin and C-peptide increased significantly. Moreover, cystatin C
and neutrophil gelatinase-associated lipocalin decreased. Collagen fibre deposition is increased with an apparent thickening of the parietal layer of Bowman’s
capsules and the basal lamina of convoluted tubules, as well as increase of the
immune-reaction of caspase-3 and desmin. The introduction of Hyphaene thebaica
led to greater amelioration in the biochemical markers, apoptotic alterations, and
podocyte injuries of the protected group than in the curative group.
Conclusions: Hyphaene thebaica may be advised as a good choice that can delay
diabetic renal complications. (Folia Morphol 2015; 74, 4: 447–457)
Key words: diabetic nephropathy, Hyphaene thebaica, podocyte injury,
apoptosis
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
2. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 411
CoQ10 is a compound which is synthesized endogenously that
is a potent lipophilic antioxidant capable of recycling and regen-
erating other antioxidants such as ascorbate and tocopherol. Also,
CoQ10 causes scavenging of free radicals, inhibition of lipid perox-
idation (Zhang et al., 2013). CoQ10 is a cofactor that plays a crucial
role in the mitochondria respiratory chain and ATP production
(Bhagavan and Chopra, 2006).
␣-Smooth muscle actin (␣-SMA) is a marker for myofibroblast-
like cells and hepatic stellate cells (HSC) (Mustafa, 2016). Vimentin
is an intermediate filament (IF) protein that is expressed frequently
in the cells of mesodermal origin as endothelial cells, that forms
an irregular network in endomysium and perimysium sheaths of
the myocardium (Heling et al., 2000). Vimentin expression occurs
during myocardial stress as heart failure (Sharov et al., 2005).
Endothelial nitric oxide synthase (eNOS) is shown in the
endothelium, within the heart, in cardiac conduction tissue and
in cardiac myocytes (Jones et al., 2004). In cardiac muscle, eNOS
arranges NO physiological action, as organizing endothelial func-
tion, platelet aggregation, vascular tone and cardiac contractility
(Pott et al., 2006). The expression of eNOS in the myocardium is
modulated in dilated cardiomyopathy with evidence of heart fail-
ure (Crespo et al., 2008). Thus, the aim of this work is to declare the
possible ameliorative potentials of CoQ10 or L-carnitine on DOX
induced cardiotoxicity.
2. Material and methods
2.1. Ethical approval
The study was conducted after approval by the Medical Research
Ethics Committee of the National Research Centre, Cairo, Egypt and
followed the recommendations of the National Institutes of Health
Guide for Care and Use of laboratory Animals (NIH Publications No.
8023, revised 1978).
2.2. Animals
The study was performed on female Wistar albino rats (n = 72),
8–10 weeks of age and weighed ranging 150–200 g that were bred
and obtained from Animal House Colony, National Research Centre,
Cairo, Egypt. All animals were housed in cages in a temperature con-
trolled (24 ± 1 ◦C) with a 12 h light/dark cycle and 60 ± 5% humidity
and were provided with standard laboratory diet and water ad libi-
tum. DOX was provided by Sigma, which was dissolved in sterile
saline. CoQ10 and L-carnitine were obtained from Mepaco, Egypt.
2.3. Experiment
Rats were divided into six groups, including 12 animals: Control
group; DOX group (10 mg/kg) (Mustafa et al., 2015); CoQ10 group
(200 mg/kg)(Mustafa et al., 2015); L-carnitine group (100 mg/kg)
(Mescka et al., 2016); DOX + CoQ10 group; DOX + L-carnitine group.
CoQ10 and L-carnitine treatment orally started 5 days before a sin-
gle dose of 10 mg/kg DOX that was injected intraperitoneally (IP)
then the treatment was continued for 10 days. At the beginning and
at the end of the study the animals body weights were measured.
2.4. Echocardiographic study
ECG was recorded at the beginning of the experiment to ensure
the normal ECG pattern of the rats. At the last day of the experiment,
rats were anesthetized by dimethyl ether and ECG was recorded for
1 min. Heart rate, P duration, QRS Interval, QTc, and ST Height were
monitored using ECG Powerlab module which consists of Power-
lab/8sp and Animal Bio-Amplifier, Australia, in addition to Lab Chart
7 software with ECG analyzer (Hajrasouliha et al., 2004).
At the end of treatment, the animals were kept for an overnight
fasting and the blood samples were collected from retroorbital
plexus and allowed to clot for 30 min at room temperature. After
blood collection, all animals were rapidly sacrificed and the hearts
were dissected and immediately homogenized in 50 mM ice-cold
phosphate buffer (pH 7.4) to give 10% homogenate (w/v). The
homogenate was centrifuged at 3200 rpm for 20 min in cooling cen-
trifuge. The supernatant was used for the determination of different
parameters.
2.5. Biochemical measurements
2.5.1. Measurement of malondialdehyde (MDA) and reduced
glutathione (GSH)
Measurement of malondialdehyde (MDA) and reduced glu-
tathione (GSH) levels using colorimetric assay kits (Catalogues No.
MD 25 29, GR 25 11 respectively) in accordance with the manufac-
turer’s instructions (Bio Diagnostic, Cairo, Egypt).
2.5.2. Measurement of Nitric oxide (NO)
Nitric oxide metabolites (NO) were determined according to
the method described by Miranda et al. (Miranda et al., 2001)
and expressed as M/g wet tissue using colorimetric assay kits
(Catalogue No. NO 25 33) in accordance with the manufacturer’s
instructions (Bio Diagnostic, Cairo, Egypt). Nitric oxide has a short
biological half-life and is rapidly converted into its stable metabo-
lites, nitrite and nitrate. Determination of nitrite and nitrate (NOx)
in body fluid and tissues is widely used as a marker of NO produc-
tion Miranda et al. (Miranda et al., 2001). Nitric oxide measured
as nitrite was determined using Griess reagent, according to the
method of Moshage et al. (Moshage et al., 1995), where nitrite, sta-
ble end product of nitric oxide radical, is mostly used as an indicator
for the production of nitric oxide.
2.5.3. Assessment of inflammatory cytokines
Serum interleukin-1beta (IL-1 ), tumor necrosis factor (TNF-
␣) and Leptin levels were measured using an enzyme-linked
immunosorbent assay (ELISA) kit (Catalogues No. RAB0277 Sigma,
RAB0479 Sigma and RAB0335 Sigma respectively) according to the
manufacturer’s instructions (Sigma-Aldrich, St. Louis, MO, United
States). All samples were tested in duplicate and averaged.
2.5.4. Assessment of cardiac markers
Lactate dehydrogenase (Catalog No. MA5-17242) and cardiac
specific creatinine kinase levels were measured using commercial
kits (Catalog No. LF-MA0233) purchased from Invitrogen (Thermo
Fisher Scientific, Inc., Waltham, MA, USA) according to the manu-
facturer’s protocol. All measurements were performed in duplicate.
2.5.5. Quantitative estimation of serum troponin I
Quantitative estimation of serum troponin I levels were car-
ried out by ELISA technique-using kit (Catalog No. LS-F127394)
purchased from Lifespan BioSciences international Inc., USA
2.5.6. Measurement of serum Troponin-T (cTnT) levels
Troponin-T (cTnT) levels were measured cTnT with a third-
generation cardio-specific assay (Catalog No. 04660307190)
(ElecsysR Troponin T STATimmunoassay manufactured by Roche
Diagnostics, France)
2.5.7. Measurement of Cardiotrophin-1
Cardiotrophin-1 is measured based on the sandwich ELISA
principle following the manufacturer’s instructions using Rat
CTF1/Cardiotrophin-1 ELISA Kit (Catalog No. LS-F127394), pur-
chased from LifeSpan BioSciences international Inc., USA.
3. 412 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Table 1
Effect of CoQ10 and L-carnitine on body weight, heart weight, heart/body weight% and mortality No.
Groups Control N = 12 CoQ10 N = 12 L-carnitine
N = 12
DOX N = 9 DOX + CoQ10
N = 11
DOX + L-
carnitine
N = 11
Body weight
(g)
152.3 ± 2.31 153.01 ± 3.41
NS
151.33 ± 5.61
NS
135 ± 6.51
1
P ≤ 0.001
146.07 ± 3.80
1
P ≤ 0.05
2
P ≤ 0.001
144.4 ± 5.03
1
P ≤ 0.01
2
P ≤ 0.001
Heart weight
(g)
0.810 ± 0.019 0.79 ± 0.021
NS
0.83 ± 0.017
NS
0.482 ± 0.040
P1
≤ 0.001
0.603 ± 0.011
1
P ≤ 0.001
2
P ≤ 0.001
0.594 ± 0.026
1
P ≤ 0.001
2
P ≤ 0.001
Heart
weight/Body
weight%
0.532 ± 0.002 0.516 ± 0.006
1
P ≤ 0.001
0.548 ± 0.008
1
P ≤ 0.001
0.357 ± 0.007
1
P ≤ 0.001
0.413 ± 0.003
1
P ≤ 0.001
2
P ≤ 0.001
0.411 ± 0.001
1
P ≤ 0.001
2
P ≤ 0.001
Mortality No. 0 0 0 3 1 1
Values are means ± SD (Control n = 12& DOX = 9 & treated = 11). ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX.
Table 2
Comparison of electrocardiographic changes in different studied groups.
Groups Heart Rate
(bpm)
P-R (s) QRS Interval (s) QTc duration
(s)
P amplitude
(mV)
T amplitude
(mV)
S-T Height
(mV)
Control
(n = 12)
318.386 ± 47.582 0.164 ± 0.017 0.015 ± 0.002 0.096 ± 0.005 0.061 ± 0.018 0.185 ± 0.043 0.035 ± 0.033
DOX 281.930 ± 22.591 0.214 ± 0.019 0.016 ± 0.004 0.134 ± 0.077 0.057 ± 0.057 0.145 ± 0.105 0.090 ± 0.066
Significance
(n = 9)
1
P = 0.013 1
P = 0.0001 1
P = 0.624 1
P = 0.024 1
P = 0.743 1
P = 0.141 1
P = 0.006
CoQ10 286.167 ± 10.913 0.187 ± 0.024 0.017 ± 0.002 0.101 ± 0.005 0.066 ± 0.007 0.174 ± 0.021 0.059 ± 0.030
Significance
(n = 12)
1
P = 0.061;
2
P = 0.815
1
P = 0.059;
2
P = 0.016
1
P = 0.275;
2
P = 0.443
1
P = 0.811;
2
P = 0.082
1
P = 0.796;
2
P = 0.625
1
P = 0.750;
2
P = 0.409
1
P = 0.405;
2
P = 0.116
L-carnitine 307.483 ± 44.887 0.182 ± 0.031 0.015 ± 0.001 0.103 ± 0.014 0.066 ± 0.009 0.209 ± 0.058 0.023 ± 0.019
Significance
(n = 12)
1
P = 0.520;
2
P = 0.161
1
P = 0.111;
2
P = 0.003
1
P = 0.698;
2
P = 0.447
1
P = 0.773;
2
P = 0.148
1
P = 0.774;
2
P = 0.604
1
P = 0.482;
2
P = 0.071
1
P = 0.335;
2
P = 0.002
DOX + CoQ10 340.989 ± 8.550 0.179 ± 0.007 0.017 ± 0.002 0.096 ± 0.011 0.047 ± 0.031 0.127 ± 0.063 0.065 ± 0.008
Significance
(n = 11)
1
P = 0.130;
2
P = 0.0001
1
P = 0.123;
2
P = 0.0001
1
P = 0.140;
2
P = 0.276
1
P = 0.975;
2
P = 0.009
1
P = 0.260;
2
P = 0.422
1
P = 0.022;
2
P = 0.486
1
P = 0.216;
2
P = 0.154
DOX + L-
carnitine
311.536 ± 36.491 0.188 ± 0.022 0.014 ± 0.003 0.105 ± 0.036 0.068 ± 0.29 0.170 ± 0.075 0.037 ± 0.035
Significance
(n = 11)
1
P = 0.560;
2
P = 0.030
1
P = 0.015;
2
P = 0.002
1
P = 0.336;
2
P = 0.119
1
P = 0.603;
2
P = 0.069
1
P = 0.588;
2
P = 0.386
1
P = 0.050;
2
P = 0.036
1
P = 0.755;
2
P = 0.004
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX.
Bpm: beat per minute. S: seconds. mV: millivolts.
2.6. Histological studies
2.6.1. Light microscopic study
Tissues were fixed in 10% neutral buffered formalin and 5 m in
thickness sections were prepared. For each specimen, at least three
to five slides were stained with H&E (hematoxylin and eosin) for
general examination, Masson’s trichrome stain to demonstrate col-
lagen fibers. Slides were observed with Olympus BX53 microscope
equipped with DP73 camera (Olympus, Tokyo, Japan) (Mustafa,
2015). The scoring system for the severity of changes was quan-
titated from none (0) to severe (4) based on the degree of necrosis,
cytoplasmic vacuolations, myocardial disorganization, degenera-
tion edema and inflammatory cell infiltrate (Alpsoy et al., 2013;
Dudka et al., 2012; Gala, 2013; Mandziuk et al., 2015).
2.6.2. Immunohistochemical study
Streptavidin–biotin peroxidase technique was applied to
paraffin-embedded tissue. 5 sections were de-waxed and pre-
treated with 3% H2O2 (hydrogen peroxide) to block endogenous
peroxidase activity. Microwave-assisted antigen retrieval was per-
formed for 10 min in 0.01 M sodium citrate buffer (pH 6.0) at
95 ◦C, and then, the slides were cooled at room temperature for
20 min. Blocking non-specific binding by incubating in 3% BSA/PBS
(Bovine Serum Albumin/Phosphate buffered saline) for 10 min.
Then, slides were incubated overnight at 4 ◦C with the primary
antibody against ␣-SMA (a mouse monoclonal antibody [Dako,
Carpinteria, California, USA] with a dilution of 1:1000; cellular
site was cytoplasmic) to evaluate the fibrosis. They were similarly
incubated with vimentin (a Mouse monoclonal antibody [Dako,
Carpinteria, California, USA] with a dilution of 1:400; cellular site
was cytoplasmic) as a cytoskeleton marker for cardiac fibroblasts
and endothelial cells and pericytes. They were incubated with
eNOS (a rabbit polyclonal antibody [Santa Cruz Biotechnology, CA,
USA] with a dilution of 1:50; cellular site was cytoplasmic) is
involved in the modulation of cardiac myocyte function. Sections
were incubated at room temperature with HRP (horseradish per-
oxidase) conjugate as a secondary antibody (Invitrogen, Zymed,
Burlington, ON, Canada). Sections were then incubated with DAB
(3,3 -diaminobenzidine tetrachloride; Vector Laboratories, Orton
Southgate, Peterborough, United Kingdom) substrate chromogen
solution (1 drop of DAB chromogen/1 mL of substrate buffer)
for 5 min to detect immunoreactivity. All sections were counter-
stained with Mayer’s hematoxylin and negative control sections
were prepared by omitting the primary antibody. While positive
control standard slides were used to prove the success of the tech-
nique. All slides were examined and the presence of labeled cells
was documented. Absence of staining was recognized as a negative
result (−), while the presence of brown staining was recognized as
positive result (+) (Mustafa, 2016).
4. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 413
Fig. 1. Effect of CoQ10 and L-carnitine on body weight, heart weight and heart/body weight%.
Values are means ± SD (Control n = 12& DOX = 9 & treated = 11). ANOVA followed by Bonferroni’s post hoc test.
1P: compared to control. 2P: compared to DOX.
Table 3
Comparison of measured oxidative stress parameters in heart tissue homogenate in different studied groups.
Groups Malondialdehyde
(nM/g)
Reduced
glutathione
(M/g)
Nitric oxide
(M/g)
Control
(n = 12)
11.79 ± 2.89 1.06 ± 0.19 10.00 ± 0.65
DOX 65.60 ± 9.46 0.82 ± 0.17 27.71 ± 8.96
Significance
(n = 9)
1
P = 0.0001 1
P = 0.005 1
P = 0.0001
CoQ10 17.18 ± 1.66 1.26 ± 0.11 10.49 ± 0.52
Significance
(n = 12)
1
P = 0.202;
2
P = 0.0001
1
P = 0.022;
2
P = 0.0001
1
P = 0.840;
2
P = 0.0001
L-carnitine 19.10 ± 2.24 1.31 ± 0.06 11.34 ± 0.59
Significance
(n = 12)
1
P = 0.101;
2
P = 0.0001
1
P = 0.004;
2
P = 0.0001
1
P = 0.579;
2
P = 0.0001
DOX + CoQ10 40.77 ± 7.44 1.15 ± 0.10 18.77 ± 1.38
Significance
(n = 11)
1
P = 0.0001;
2
P = 0.0001
1
P = 0.266;
2
P = 0.0001
3
P = 0.001;
2
P = 0.001
DOX + L-
carnitine
41.03 ± 10.73 1.08 ± 0.08 16.29 ± 1.51
Significance
(n = 11)
1
P = 0.0001;
2
P = 0.0001
1
P = 0.840;
2
P = 0.003;
1
P = 0.015;
2
P = 0.0001
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX. nM/g: nanomolar/gram. M/g: micromolar/gram.
2.6.3. Morphometric study
Ten non-overlapping fields for each animal were selected ran-
domly and analyzed to determine cardiomyocytes’ diameter of
H&E stained sections. Cardiomyocytes with centrally located visible
nuclei intact cell membrane were selected and the measurements
were done along their short axis (de Salvi Guimaraes et al., 2017;
Nascimento et al., 2016; Pradegan et al., 2016). The area percent-
age of collagen fibers in Masson’s trichrome, ␣-SMA, vimentin and
eNOS-stained sections. Quantitative measurements were analyzed
with the use of Image-Pro Plus v6 (Media Cybernetics Inc., Bethesda,
Maryland, USA) and ImageJ (NIH, 1.51; Melville, NY, USA), which
was calibrated for distance, color and area before its use (Mustafa
and Hussein, 2015).
2.6.4. Ultrastructure study
One mm3 samples were immersed in 2.5% glutaraldehyde in
0.1 M phosphate buffer at 4 ◦C for 3 hs and post-fixed in 1% OsO4
(osmium tetraoxide). Then, tissues were embedded in Epon 812 and
semithin sections were prepared, stained with toluidine blue and
observed with a microscope. Ultrathin sections of 50–60 nm thick
5. 414 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Fig. 2. A: Comparison of electrocardiographic changes in different studied groups [Heart Rate (bpm)]. B: Comparison of electrocardiographic changes in different stud-
ied groups [P-R duration (seconds)]. C: Comparison of electrocardiographic changes in different studied groups [QRS Interval (seconds)]. D: Comparison of electrocardiographic
6. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 415
Table 4
Comparison of measured inflammatory parameters in different studied groups.
Groups Interleukin-1
(pg/ml)
Tumor necrosis
factor-␣
(pg/ml)
Leptin (pg/ml) Lactate
dehydrogenase
(U/ml)
Control
(n = 12)
40.35 ± 7.47 40.25 ± 4.48 27.12 ± 3.45 100.80 ± 12.24
DOX 286.01 ± 24.55 198.00 ± 12.43 120.14 ± 10.51 403.40 ± 37.83
Significance
(n = 9)
1
P = 0.0001 1
P = 0.0001 1
P = 0.0001 1
P = 0.0001
CoQ10 45.43 ± 8.82 53.86 ± 22.80 27.54 ± 6.86 96.20 ± 18.10
Significance
(n = 12)
1
P = 0.597;
2
P = 0.0001
1
P = 0.077;
2
P = 0.0001
1
P = 0.932;
2
P = 0.0001
1
P = 0.740;
2
P = 0.0001
L-carnitine 42.28 ± 5.27 37.19 ± 3.93 27.36 ± 7.19 105.00 ± 17.92
Significance
(n = 12)
1
P = 0.840;
2
P = 0.0001
1
P = 0.682;
2
P = 0.0001
1
P = 0.961;
2
P = 0.0001
1
P = 0.761;
2
P = 0.0001
DOX + CoQ10 84.44 ± 8.82 72.24 ± 6.52 48.66 ± 6.35 131.60 ± 18.37
Significance
(n = 11)
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.0001
1
P = 0.034;
2
P = 0.0001
DOX + L-
carnitine
143.51 ± 21.67 113.61 ± 7.90 84.72 ± 9.80 182.80 ± 15.51
Significance
(n = 11)
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.0001
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX.
pg/ml: Picograms per Millilitre. U/ml: Units per Millilitre.
Table 5
Comparison of measured heart parameters in different studied groups.
Groups Cardiotrophin-
1
(pg/ml)
Cardiac
specific-
creatine kinase
(ng/ml)
Troponin-I
(ng/ml)
Troponin-T
(ng/ml)
Control
(n = 12)
68.20 ± 8.47 100.80 ± 12.24 0.72 ± 0.06 0.39 ± 0.08
DOX 237.36 ± 18.01 403.40 ± 37.83 5.80 ± 0.74 1.81 ± 0.55
Significance
(n = 9)
1
P = 0.0001 1
P = 0.0001 1
P = 0.0001 1
P = 0.0001
CoQ10 62.45 ± 6.19 96.20 ± 18.10 0.70 ± 0.09 0.43 ± 0.07
Significance
(n = 12)
1
P = 0. 0.375;
2
P = 0.0001
1
P = 0.740;
2
P = 0.0001
1
P = 0.952;
2
P = 0.0001
1
P = 0.850;
2
P = 0.0001
L-carnitine 64.08 ± 6.22 105.00 ± 17.92 0.68 ± 0.15 0.44 ± 0.13
Significance
(n = 12)
1
P = 0. 0.523;
2
P = 0.0001
1
P = 0.761;
2
P = 0.0001
1
P = 0.886;
2
P = 0.0001
1
P = 0.792;
2
P = 0.0001
DOX + COQ10 109.78 ± 9.10 131.60 ± 18.37 1.44 ± 0.38 0.98 ± 0.33
Significance
(n = 11)
1
P = 0.0001;
2
P = 0.0001
1
P = 0.034;
2
P = 0.0001
1
P = 0.011;
2
P = 0.0001
1
P = 0.004;
2
P = 0.0001
DOX + L-
carnitine
167.08 ± 7.05 182.80 ± 15.51 2.37 ± 0.55 1.39 ± 08
Significance
(n = 11)
1
P = 0.0001;
2
P = 0.001
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.0001
1
P = 0.0001;
2
P = 0.23
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX. pg/ml: picograms per Millilitre. ng/ml: Nanograms per Millilitre.
were cut by ultramicrotome (NOVA, LKB 2188, Bromma, Sweden);
and stained with uranyl acetate and lead citrate. Then tissues
were examined with Philips 201 transmission electron micro-
scope (Philips Industries, Eindhoven, Netherlands) at 60–80 kv at
the Transmission Electron Microscope Unit (Mustafa and Hussein,
2015).
2.7. Statistical analysis
Statistical Analysis. Quantitative data were expressed as the
mean and standard deviations. Data were analyzed using a one-
way analysis of variance (ANOVA) followed by Bonferroni’s post
hoc test. All statistical analyses were implemented using the Sta-
tistical Package for the Social Sciences (SPSS), version 23. The values
were considered significant when P < 0.05 (Mustafa, 2015).
3. Results
3.1. General assessment
The results revealed a significant decrease in the heart/body
weight ratio in DOX group. The administration of CoQ10 or
L-carnitine significant increase in the heart/body weight ratio
(Table 1, Fig. 1).
changes in different studied groups [T amplitude (mV)]. E: Comparison of electrocardiographic changes in different studied groups [S-T Height (mV)]. (F&G): ECG of control
group. ECG of DOX group. Rats were anesthetized and ECG was recorded for 1 min. PVC: premature ventricular complex. AV block: Atrio-ventricular block.
7. 416 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
3.2. ECG findings
Heart rate was significantly lower in DOX group than control,
DOX + CoQ10 and DOX + L-carnitine groups. P-R duration was sig-
nificantly higher in DOX + L-carnitine group versus control and was
significantly higher in DOX group than control, CoQ10, L-carnitine,
DOX + CoQ10 and DOX + L-carnitine groups. QTc was significantly
higher in DOX group than control and DOX + CoQ10. T amplitude
was significantly lower in DOX group than DOX + L-carnitine. S-T
height was significantly lower in DOX group than control, L-
carnitine and DOX + L-carnitine (Table 2, Fig. 2A–I).
3.3. Heart tissue homogenate levels of oxidative stress markers
Heart tissue homogenate levels of MDA was significantly higher
in DOX group than control, CoQ10, L-carnitine, DOX + CoQ10
and DOX + L-carnitine groups and in DOX + CoQ10 and DOX + L-
carnitine groups versus control. Heart tissue homogenate levels
of NO was significantly higher in DOX group than control, CoQ10,
L-carnitine, DOX + CoQ10 and DOX + L-carnitine groups and in
DOX + CoQ10 and DOX + L-carnitine groups versus control. Heart
tissue homogenate levels of reduced glutathione was significantly
lower in DOX group than control, CoQ10, L-carnitine, DOX + CoQ10
and DOX + L-carnitine groups but was significantly higher in CoQ10
and L-carnitine groups versus control (Table 3, Fig. 3A–C).
3.4. Serum levels of inflammatory cytokines
Serum levels of IL-1 beta, TNF-␣ and leptin were significantly
higher in DOX group than control, CoQ10, L-carnitine, DOX + CoQ10
and DOX + L-carnitine groups and in DOX + CoQ10 and DOX and
L-carnitine groups versus control. Serum level of LDH was sig-
nificantly higher in DOX group than control, CoQ10, L-carnitine,
DOX + CoQ10 and DOX + L-carnitine groups and in DOX + CoQ10 and
DOX + L-carnitine groups versus control (Table 4, Fig. 4A–D).
3.5. Serum levels of cardiac parameters
Serum levels of Cardiotrophin-1, Cardiac specific-creatine
kinase, and Troponin-I were significantly lower in control, CoQ10,
L-carnitine, DOX +CoQ10 and DOX + L-carnitine than DOX group
while regarding Troponin-T no significant difference between DOX
group and DOX + L-carnitine group. Serum levels of Cardiotrophin-
1, Cardiac specific-creatine kinase, Troponin-I and Troponin-T in
DOX + CoQ10 and DOX + L-carnitine groups versus control showed
a significant increase (Table 5, Fig. 5A–D).
3.6. Histological studies
Control group. H&E stained sections of control heart tissues
showed normal cardiac myocytes with their centrally placed nuclei
(Fig. 6A). Sections stained with Masson’s trichrome stain showed
scanty green stained connective tissue surrounding the muscle
fibers (Fig. 7A). Groups treated with only L-carnitine and CoQ10
revealed no significant differences between these groups and con-
trol as regard H&E and Masson’s trichrome stains.
DOX group. H&E showed necrosis and swollen of the car-
diomyocytes with an increase in the diameter. Pyknotic nuclei,
mononuclear cellular infiltration and dilated blood vessels were
observed (Fig. 6B). Masson’s trichrome stained sections showed
intense increase in collagen fibers of the surrounding endomysium
(Fig. 7B). These results were confirmed by morphometric and sta-
tistical study. Cardiomyocytes diameter of DOX group showed a
significant decrease in the mean cardiomyocyte diameter (P < 0.01)
when compared with the control. Area percentage of collagen (Mas-
son’s trichrome stain) of DOX group showed a significant increase
in the area percentage of collagen (P < 0.001) when compared with
the control (Tables 6, 7).
DOX + CoQ10 group: H&E showed nearly normal microscopic
architecture of cardiomyocytes with minimal changes in nuclei
were observed (Fig. 6C). Masson’s trichrome stained sections
showed mild reaction (Fig. 7C). These results were confirmed
by morphometric and statistical study (Tables 6, 7). DOX + L-
carnitine group: H&E showed apparently normal microscopic
histo-architecture of cardiomyocytes with mild changes in nuclei
were observed (Fig. 6D). Masson’s trichrome stained sections
showed mild reaction (Fig. 7D). These results were confirmed by
morphometric and statistical study (Tables 6, 7). Cardiomyocytes
diameter of groups treated with CoQ10 and L-carnitine showed a
significant improvement as compared with DOX group (Table 6,
Fig. 11). Area percentage of collagen (Masson’s trichrome stain) of
groups treated with CoQ10 and L-carnitine showed a significant
improvement as compared with the DOX group (Tables 6, 7 Fig. 11).
3.6.1. Immunohistochemical results for ˛-SMA
Immunohistochemical Results for ␣-SMA of the control group
revealed minimal immune expression (Fig. 8A). DOX group
showedextensiveimmuneexpression(Fig.8B).DOX + CoQ10group
revealed mild immune expression (Fig. 8C). DOX + L-carnitine
group showed moderate immune expression (Fig. 8D). Mean area%
of ␣–SMA immunopositive cells of DOX group showed a signifi-
cant increase in the mean area% of ␣–SMA immunoreactivity when
compared with the control. Also, groups treated with CoQ10 and
L-carnitine showed a significant improvement as compared with
the DOX group (Table 7, Fig. 11).
3.6.2. Immunohistochemical results for vimentin
Immunohistochemical Results for vimentin of the control
group revealed faint immune expression in the myocardium
(Fig. 9A). DOX group showed wide positive immunoreactivity in
the myofibroblasts (Fig. 9B). DOX + CoQ10 group revealed mini-
mal immune expression (Fig. 9C). DOX + L-carnitine group showed
slight immune expression (Fig. 9D). Mean area% of vimentin of DOX
group showed a significant increase in the mean area% of vimentin
immunoreactivity when compared with the control. In addition,
groups treated with CoQ10 and L-carnitine showed a significant
improvement as compared with the DOX group (Table 7, Fig. 11).
3.6.3. Immunohistochemical Results for vimentin
Immunohistochemical Results for eNOS of the control group
revealed faint or no immune expression (Fig. 10A). DOX group
showed strong immune expression (Fig. 10B, C). DOX + CoQ10
group revealed minimal immune expression (Fig. 10D). DOX + L-
carnitine group showed moderate immune expression (Fig. 10E).
Mean area% of eNOS of DOX group showed a significant increase
in the mean area% of eNOS immunoreactivity when compared
with the control. Also, groups treated with CoQ10 and L-carnitine
showed a significant improvement as compared with the DOX
group (Table 7, Fig. 11).
3.6.4. Immunohistochemical Results for eNOS
Ultrastructural results. Control group showed normal architec-
ture of the cardiomyocytes (Figs. 12 and 13A). DOX group revealed
degeneration and fragmentation of myofibrils and loss of light
bands with broadening and interruption of Z lines. The mitochon-
dria appeared electron dense with a moth eaten appearance among
the muscle fibers (Figs. 12 and 13B). DOX + CoQ10 group revealed
well-organized myofibrils and the mitochondria looked normal
with tightly packed cristae (Figs. 12 and 13C). DOX + L-carnitine
showed an improvement of the myofibrils organization (Figs. 12
and 13D).
8. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 417
Fig. 3. A: Comparison of measured oxidative stress parameters in heart tissue homogenate in different studied groups for Malondialdehyde (nM/g). B: Comparison of
measured oxidative stress parameters in heart tissue homogenate in different studied groups for reduced glutathione (M/g). C: Comparison of measured oxidative stress
parameters in heart tissue homogenate in different studied groups for Nitric oxide (M/g).
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX. nM/g: nanomolar/gram.
9. 418 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Fig. 4. A: Comparison of measured inflammatory parameters in different studied groups [Interleukin-1 (pg/ml)]. B: Comparison of measured inflammatory parameters in
different studied groups [Tumor necrosis factor-␣ (pg/ml)]. C: Comparison of measured inflammatory parameters in different studied groups [Leptin (pg/ml)]. D: Comparison
of measured inflammatory parameters in different studied groups [Lactate dehydrogenase (U/ml)].
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX.
pg/ml: Picograms per Millilitre.
Fig. 5. A: Comparison of measured heart parameters in different studied groups [Cardiotrophin-1 (pg/ml)]. B: Comparison of measured heart parameters in different studied
groups [Cardiac specific-creatine kinase (ng/ml)]. C: Comparison of measured heart parameters in different studied groups [Troponin-I (ng/ml)]. D: Comparison of measured
heart parameters in different studied groups [Troponin-T (ng/ml)].
Values are means ± SD. ANOVA followed by Bonferroni’s post hoc test.
1
P: compared to control. 2
P: compared to DOX. pg/ml: picograms per Millilitre.
10. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 419
Fig. 6. (A). Photomicrograph of control showed cardiac myocytes with centrally placed nuclei (arrow). (B). DOX treated group showed cardiac myocytes showing massive
necrosis with focal marked fragmentation and nuclear changes in the form of pyknosis (p), karyolysis (k) and chromatin margination (c). (C). CoQ10 and DOX showed nearly
normal architecture of the cardiac myocytes with focal necrosis. (D): L-carnitine and DOX showed apparently regular architecture of the cardiac myocytes with focal necrosis
(H&E, Scale bar 20 m).
Fig. 7. (A). Photomicrograph of control showed scanty green colored collagen fibers (arrow) between the cardiomyocytes. (B). DOX treated group showed an intense of
greenish colored collagen fibers (arrow) between swollen cardiomyocytes. (C). CoQ10 and DOX showed mild reaction. (D): L-carnitine and DOX showed mild reaction
(Masson’s trichrome, Scale bar 20 m). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.).
4. Discussion
The cardiotoxicity of doxorubicin (DOX) limits its use in cancer
chemotherapy; the cells that are most affected by DOX are those
with a large number of mitochondria, which include cardiac and
liver cells. New approaches are therefore needed to decrease the
oxidative side effects of doxorubicin (Chao et al., 2011). DOX pos-
sesses cardiotoxic properties that affect both the conductivity and
11. 420 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Fig. 8. (A). Photomicrograph of control showed faint immunoreactivity in the myocardium (arrow). (B). DOX treated group showed wide positive immunoreactivity in the
myofibroblasts, which are attached together by their processes (arrow). (C). CoQ10 and DOX showed minimal immunoreactivity. (D). L-carnitine and DOX showed slight
immunoreactivity (arrow) (arrow) (␣-SMA. Scale bar 20 m).
Fig. 9. (A) Photomicrograph of control showed minimal immune reaction in the blood capillaries wall (curved arrows) and interstitial cells (arrow). With an immune negative
cardiac muscle fibers (arrowhead). (B). DOX treated group showed strong immune reaction in endomysium and perimysium connective tissues (star), in the blood capillaries
wall (curved arrows), and interstitial cells (arrow). (C). CoQ10 and DOX showed mild immune reaction in the endomysium and perimysium (star), in the blood capillaries wall
(curved arrows) and interstitial cells (arrow). With an immune negative reaction in cardiac muscle fibers (arrowhead). (D): L-carnitine and DOX showed moderate immune
reaction (Vimentin. Scale bar 20 m).
12. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 421
Fig. 10. (A). Photomicrograph of control showed faint or no immune reaction cardiac muscle fibers. (B, C). DOX treated group showed strong immune reaction (arrowhead)
in cardiac muscle fibers and endothelial cells of blood capillaries. (C). CoQ10 and DOX showed minimal immune reaction (arrowhead). (E): L-carnitine and DOX showed
moderate immune reaction (arrowhead) (eNOS. Scale bar 5 m).
Table 6
Effect of CoQ10and L-carnitine on the heart tissues treated with DOX.
Groups Control DOX DOX + CoQ10 DOX + L-Carnitine
Necrosis 0*
+4 +1 +1
Degeneration and vacuolations 0 +4 +1 +1
Edema 0 +3 +1 +1
Inflammatory cell infiltrate 0**
+3 +1 +1
A single animal may be represented more than once in the listing of individual histological changes. *Massive necrosis/changes limited to single cardiomyocytes. **Massive
inflammatory infiltration/disseminate mononuclear cells between cadiomyocytes.
Table 7
Cardiomyocyte diameter, area percentage of collagen, vimentin, ␣-SMA and eNOS Immunohistochemistry of the different groups.
Groups Control N = 12 CoQ10 N = 12 L-Carnitine
N = 12
DOX N = 9 DOX + CoQ10
N = 11
DOX + L-
Carnitine
N = 11
Cardiomyocyte
diameter (m)
14.24 ± 2.71 15.35 ± 3.07 13.84 ± 2.3 10.01 ± 0.97
1
P ≤ 0.001
16.92 ± 1.082
1
P ≤ 0.01
2
P ≤ 0.001
16.25 ± 1.071
1
P ≤ 0.05
2
P ≤ 0.001
Area
percentage of
collagen (m2
)
4.57 ± 1.52 3.82 ± 1.27 5.32 ± 1.77 23.17 ± 3.61
1
P ≤ 0.001
9.01 ± 0.70
1
P ≤ 0.001
2
P ≤ 0.001
11.72 ± 1.41
1
P ≤ 0.001
2
P ≤ 0.001
Area
percentage of
␣-SMA
0.16 ± 0.05 0.91 ± 0.3 0.66 ± 0.22 12.67 ± 1.97
1
P ≤ 0.001
5.12 ± 1.36
1
P ≤ 0.001
2
P ≤ 0.001
6.32 ± 1.14
1
P ≤ 0.001
2
P ≤ 0.001
Area
percentage of
vimentin
0.37 ± 0.12 1.12 ± 0.37 0.87 ± 0.29 31.21 ± 4.45
1
P ≤ 0.001
10.02 ± 2.13
1
P ≤ 0.001
2
P ≤ 0.001
9.97 ± 1.08
1
P ≤ 0.001
2
P ≤ 0.001
Area
percentage of
eNOS
1.23 ± 0.36 1.48 ± 0.49 0.98 ± 0.33 6.45 ± 2.15
1
P ≤ 0.001
1.97 ± 0.32
1
P = NS
2
P ≤ 0.001
1.64 ± 0.54
1
P = NS
2
P ≤ 0.001
Values are means ± SD (Control n = 12& DOX = 9 & treated = 11). ANOVA followed by Bonferroni’s post hoc test.
1P: compared to control. 2P: compared to DOX.
rhythmicity of cardiac muscle, as shown by its effect on heart rate
in addition to the associated elongation of the corrected QT interval
(QTc), ST elevation, and shortening of the T amplitude (Mantawy
et al., 2014).
The results of this study showed significant abnormalities that
affected ECG in the DOX group in agreement with previous studies
(Goyal et al., 2016; Jagetia and Venkatesh, 2015). These changes
include reflected arrhythmias, conduction abnormalities, and the
attenuation of left ventricular function (Mantawy et al., 2014).
This study illustrated that DOX induces oxidative damage and
nitrosative stress in the cardiac muscle. These results align with
those from other studies (Goyal et al., 2016; Jagetia and Venkatesh,
2015). These results could be explained by the ability of DOX to gen-
erate ROS, which results in lipid peroxidation of both the cellular
and mitochondrial membrane, ending in the injury of myocardio-
13. 422 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Fig. 11. Cardiomyocyte diameter and Area% of collagen,␣-SMA, vimentin and eNOS. The mean is given in columns, and error bars represent the standard deviation (SD).
cytes (Sahu et al., 2016). Moreover, DOX creates free radicals that
cause destruction in DNA and proteins and interfere with the struc-
ture of the cytoskeleton (Ikeda et al., 2010). Oxidative stress could
injure mitochondrial cell membranes, increasing the membrane’s
permeability and making it vulnerable to rupture (Viswanatha
Swamy et al., 2011).
L-carnitine produces its antioxidant effects through different
mechanisms, including the scavenging of free radical activity either
directly or by inhibition of its production, maintaining the effi-
ciency of the mitochondrial electron transport chain, stimulating
the activation of antioxidant enzymes, and synthesis of antioxidant
molecules like reduced glutathione (Surai, 2015).
L-carnitine protects myocardial integrity by controlling the
intra-mitochondrial percentage of acyl-CoA/CoA, resulting in elim-
ination of toxic compounds; maintaining the integrity of the
mitochondrial membrane’s permeability; and promoting the elim-
ination of free radicals (Chao et al., 2011).
CoQ10 plays an important role in energy metabolism and is
part of the electron transport chain that is responsible for ATP
synthesis. Moreover, it is one of the most efficient endogenous
antioxidants and protects cellular DNA, lipids, and protein from
oxidative damage (Garrido-Maraver et al., 2014). CoQ10 protects
myocardial integrity through many mechanisms, including preser-
vation of myocardial ATP levels and powerful antioxidant effects.
CoQ10 may exert its effects directly by acting as a scavenger of free
radicals or through the regeneration of tocopherol and ascorbic acid
from their oxidized state (Chen et al., 2017).
The results of this study confirm that DOX toxicity has specific
inflammatory effects, as evidenced by the significant increase in
14. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 423
Fig. 12. (A). Electron micrograph of control showed a cardiomyocyte with an elongated nucleus (N) with an evenly dispersed chromatin pattern and regular nuclear membrane
(↑). Numerous mitochondria (M) appear with apparent cristae between the longitudinally arranged myofibrils. That exhibit a normal cross-striated pattern Z lines (Z). (B).
DOX treated group showed disorganized, fragmented, degenerated myofibrils with loss of cross striations (↑). Distorted mitochondria (M) with dense matrix, unapparent
cristea, with different shapes and sizes irregularly arranged between the myofibrils and wide intercellular spaces (star) in the sarcoplasm of the cardiac myocytes. (C). CoQ10
and DOX showed regularly arranged myofilaments between successive Z lines (Z) in the sarcomeres. Mitochondria (M) arranged in rows between the myofibrils. The nucleus
(N) of a cardiac muscle fiber with slightly irregular nuclear membrane (↑). (D): L-carnitine and DOX showed mitochondria (M) appear distorted, with different shapes and
sizes around the nucleus (N) and between the myofibrils. Nuclear membrane indentations is observed (↑). Note the wide intercellular space (*) between adjacent muscle
fibers. (Scale bar 2 m).
inflammatory cytokines. These results are in agreement with other
studies (Elsherbiny et al., 2016; Sun et al., 2016).
These results might be explained by the fact that ROS produced
by DOX can initiate inflammatory responses, mainly via NF-B,
which results in the release of cytokines such as tumor necrosis
factor-alpha [TNF-␣] and interleukin 1 beta [IL-1 ] (Sun et al.,
2016).
Leptin is considered one of acute response markers in oxida-
tive stress; it is involved in the prediction of coronary heart disease
due to the known relation between C-reactive protein and leptin
(Ahmed et al., 2005). In addition, this study showed that DOX led
to significant myocardial damage, as evidenced by increased serum
levels of both CK-MB and LDH. These results, in accordance with
those of other studies (El-Agamy et al., 2016; Sun et al., 2016), can
be explained by the increase in oxidative stress leading to lipid per-
oxidation and disruption of the cell membranes of myocardiocytes,
along with the release of biochemical markers in the serum and
plasma. CK-MB is one of the most important biochemical diagnostic
markers for myocardial damage (El-Agamy et al., 2016).
Treatment with CoQ10 and L-carnitine resulted in a sig-
nificant decrease of these enzymes that is attributable to a
decrease in oxidative stress and stabilization of cardiomyocyte
cell membranes. Furthermore, specific cardiac markers for acute
cardiotoxicity have been measured, including cardiac troponin
I (cTnL), T (cTnT), and cardiotrophin-1. All of these parameters
showed significant elevation in the group treated with DOX (Atas
et al., 2015). These results are concordant with those from other
studies (Atas et al., 2015; Bertinchant et al., 2003; Reagan et al.,
2013) showing an increase in the levels of cTnI and cTnT, confirm-
ing that these are sensitive and specific markers for cardiac injury
that may be elevated in the blood of patients treated with DOX
before cardiac damage is evident. Therefore, these markers can
be used for the prediction of future left ventricular dysfunction.
They are used in early detection of necrosis, before CK-MB levels
significantly increase in the heart (Atas et al., 2015).
In the current study, the DOX group showed visible conges-
tion in between cardiomyocytes, a finding that coincided with
other findings that noted the presence of marked blood cells
in the peri-capillary space (Hadi et al., 2012). The vacuoles are
ascribed to the expansion of cytoplasmic membranous compo-
nents due to redistribution of the intra-cellular electrolytes and
water (Balli et al., 2004). The diameter of cardiomyocytes was
increased, in agreement with other reports in which myocytic
diameter increased, with the presence of hyperchromatic nuclei,
disorganization of myofibrils, and loss of cross-striation of car-
diac myocytes (Rashikh et al., 2011). With CoQ10 or L-carnitine,
15. 424 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Fig. 13. (A). Electron micrograph of control showed a cardiomyocyte contains strands of myofibrils formed of light bands (I), Z lines (Z), dark bands (A), H zone (H), sarcomere
(S) and mitochondrial rows (M). (B). DOX treated group showed destruction, fragmentation and lysis of myofibrils (arrows) with absence of light bands and broadening of Z
lines (Z). Moth-eaten appearance of degenerated mitochondria (ME) with variable sizes were seen among the myofibrils. Note lipofuscin pigment (star). (C). CoQ10 and DOX
showed myofibrils with preserved cross-banding pattern, intercalated disc (IC) and euchromatic nucleus (N). The mitochondria (m) looks normal with tightly packed cristae
and relative increase in number. (D): L-carnitine and DOX showed well-organized myofibrils with few interrupted Z lines (arrow. Preserved healthy mitochondria (M) and
Dilated SER (SER). (Scale bar 500 nm).
the histopathological findings were improved such that they were
consistent with other studies that found decreases in myofibril dis-
organization, exudation, and inflammatory cell infiltration in the
myocardium (Kwong et al., 2002). Analysis of the ultrastructure
morphology images showed peripheral chromatin condensation,
deformity and fragmentation of the nuclei, and apoptosis (Zhang
et al., 2012), supporting the hypothesis that apoptosis is one of
the mechanisms of DOX-cardiotoxicity, DOX-induced lipid per-
oxidation, reactive oxygen species (ROS) production, disturbed
mitochondrial metabolism, and direct cardiotoxicity (Oktem et al.,
2012).
The results were in agreement with those of previous
researchers who observed that myocardial stress increases the
mean number of ␣-SMA positive myofibroblasts. This was
attributed to myofibroblasts, which are considered the key cells
responsible for extracellular matrix and collagen deposition in
myocardial fibrosis (Naugle et al., 2006). Other researchers have
observed a rise in fibronectin and in collagen types I and III, ascrib-
ing this to collagen synthesis related to -adrenergic receptor
activation in fibroblasts (Yin et al., 2009). The contractile fibers of
myofibroblasts contain ␣-SMA and are linked to exaggerated extra-
cellular matrix accumulation in pathological disorders (Ma et al.,
2014). In cardiac disease, cardiomyocytes are wasted due to necro-
sis, and myofibroblasts are stimulated to launch restorative fibrosis.
Myofibroblasts also generate angiotensin II and fibrogenic growth
factors, which play a crucial role in fibrosis and collagen type I
synthesis (Weber et al., 2013). With CoQ10 or L-carnitine, there
is a decrease in the transformation of fibroblasts to myofibroblasts,
which are a source of collagen, thus restraining cardiac fibrosis.
In the current work, an increase in vimentin area percentage
expressed in the arterial walls was observed in the DOX group;
similar results were revealed in dilated cardiomyopathy, where
vimentin immunoreaction was increased in the interstitial tis-
sue cells (Di et al., 2000). The increased vimentin was linked
to an increase of collagen and fibrosis (Schaper et al., 1991).
Scientists revealed an adverse connection between myocardial
vimentin overexpression and the sliding rate of actin myosin. It
was proposed that disarrangement of cytoskeleton proteins occurs
with participation of vimentin in the modulations of coupling of
myocytes to the extracellular matrix, myocyte functions, and intra-
cellular signaling during cardiac failure and hypertrophy (Rastogi
et al., 2008). Moreover, investigators noticed the proliferation of
T-tubules linked to vimentin overexpression in cardiomyopathy.
This can lead to recovery of an inappropriate cardiac function by
substituting for the contractile elements (Di et al., 2000). Fibrosis
is responsible for an increased stiffness and decrease of ventricular
compliance (Heling et al., 2000).
16. H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426 425
The interaction between DOX and NOS is a complex. DOX
converts eNOS from a nitric oxide donor to a superoxide gen-
erator (Octavia et al., 2012). DOX-induced hydrogen peroxide
creation (H2O2) is responsible for apoptotic cell death and DOX-
toxicity (Octavia et al., 2012). In turn, H2O2 promotes endothelial
nitric oxide synthase (eNOS) transcription in endothelial cells and
cardiomyocytes (Kalyanaraman et al., 2002). Up-regulated eNOS
expression can play a key role in DOX-cardiac dysfunction by
affecting ROS-mediated apoptosis of endothelial cells (Neilan et al.,
2007). Genetic disruption of eNOS transcription protects against
DOX-induced cardiotoxicity and mortality, while overexpression
exaggerates the toxic effects of DOX (ˇSim ˚unek et al., 2009). Studies
about endothelial dysfunction have demonstrated a considerable
attenuation of endothelial vasodilation after DOX administration,
suggesting dysfunctional eNOS activity (Olukman et al., 2009).
The current results provide proof that CoQ10 and L-carnitine
attenuates DOX-induced generation of free radicals. Also, prevent
eNOS uncoupling by reducing superoxide formation, increasing
NO bioavailability, and inhibiting upregulation of the activity and
expression of the vascular NAD (P) H oxidase (Chatterjee et al.,
2010).
5. Conclusion
Supplementation with CoQ10 or L-carnitine defends the
myocardium through their antioxidant activity, as was proven by
the improvement of different biochemical markers and oxidative
status and the restoration of the myocardium’s structural integrity
and function.
Disclosure of interest
The authors declare that they have no conflicts of interest.
References
˙Izgüt-Uysal, V.N., A˘gac¸ , A., Derin, N., 2003. Effect of L-Carnitine on
carrageenan-induced inflammation in aged rats. Gerontology 49, 287–292.
ˇSim ˚unek, T., ˇStˇerba, M., Popelová, O., Adamcová, M., Hrdina, R., Gerˇsl, V., 2009.
Anthracycline-induced cardiotoxicity: overview of studies examining the roles
of oxidative stress and free cellular iron. Pharmacol. Rep. 61, 154–171.
Agarwal, A., Said, T.M., 2004. Carnitines and male infertility. Reprod Biomed Online
8, 376–384.
Ahmed, H.H., Mannaa, F., Elmegeed, G.A., Doss, S.H., 2005. Cardioprotective activity
of melatonin and its novel synthesized derivatives on doxorubicin-induced
cardiotoxicity. Bioorg. Med. Chem. 13, 1847–1857.
Aliev, G., Liu, J., Shenk, J.C., Fischbach, K., Pacheco, G.J., Chen, S.G., Obrenovich, M.E.,
Ward, W.F., Richardson, A.G., Smith, M.A., 2009. Neuronal mitochondrial
amelioration by feeding acetyl-L-carnitine and lipoic acid to aged rats. J. Cell.
Mol. Med. 13, 320–333.
Alpsoy, S., Aktas, C., Uygur, R., Topcu, B., Kanter, M., Erboga, M., Karakaya, O.,
Gedikbasi, A., 2013. Antioxidant and anti-apoptotic effects of onion (Allium
cepa) extract on doxorubicin-induced cardiotoxicity in rats. J. Appl. Toxicol. 33,
202–208.
Arcamone, F., Cassinelli, G., Fantini, G., Grein, A., Orezzi, P., Pol, C., Spalla, C., 2000.
Adriamycin, 14-hydroxydaunomycin, a new antitumor antibiotic from S.
peucetius var. caesius. reprinted from biotechnology and bioengineering, vol.
XI, issue 6, pages 1101–1110 (1969). Biotechnol. Bioeng. 67, 704–713.
Atas, E., Kismet, E., Kesik, V., Karaoglu, B., Aydemir, G., Korkmazer, N., Demirkaya,
E., Karslioglu, Y., Yurttutan, N., Unay, B., Koseoglu, V., Gokcay, E., 2015. Cardiac
troponin-I, brain natriuretic peptide and endothelin-1 levels in a rat model of
doxorubicin-induced cardiac injury. J. Cancer Res. Ther. 11, 882–886.
Balli, E., Mete, U.O., Tuli, A., Tap, O., Kaya, M., 2004. Effect of melatonin on the
cardiotoxicity of doxorubicin. Histol. Histopathol. 19, 1101–1108.
Bertinchant, J.P., Polge, A., Juan, J.M., Oliva-Lauraire, M.C., Giuliani, I.,
Marty-Double, C., Burdy, J.Y., Fabbro-Peray, P., Laprade, M., Bali, J.P., Granier, C.,
de la Coussaye, J.E., Dauzat, M., 2003. Evaluation of cardiac troponin I and T
levels as markers of myocardial damage in doxorubicin-induced
cardiomyopathy rats, and their relationship with echocardiographic and
histological findings. Clin. Chim. Acta 329, 39–51.
Bhagavan, H.N., Chopra, R.K., 2006. Coenzyme Q10: absorption, tissue uptake,
metabolism and pharmacokinetics. Free Radic. Res. 40, 445–453.
Chao, H.-H., Liu, J.-C., Hong, H.-J., Lin J.-w. Chen, C.-H., Cheng, T.-H., 2011.
L-carnitine reduces doxorubicin-induced apoptosis through a
prostacyclin-mediated pathway in neonatal rat cardiomyocytes. Int. J. Cardiol.
146, 145–152.
Chatterjee, K., Zhang, J., Honbo, N., Karliner, J.S., 2010. Doxorubicin
cardiomyopathy. Cardiology 115, 155–162.
Chen, P.-Y., Hou, C.-W., Shibu, M.A., Day, C.H., Pai, P., Liu, Z.-R., Lin, T.-Y.,
Viswanadha, V.P., Kuo, C.-H., Huang, C.-Y., 2017. Protective effect of Co-enzyme
Q10 On doxorubicin-induced cardiomyopathy of rat hearts. Environ. Toxicol.
32, 679–689.
Crespo, M.J., Zalacain, J., Dunbar, D.C., Cruz, N., Arocho, L., 2008. Cardiac oxidative
stress is elevated at the onset of dilated cardiomyopathy in
streptozotocin-diabetic rats. J. Cardiovasc. Pharmacol. Ther. 13, 64–71.
Di, S., Marotta, M., Salvatore, G., Cudemo, G., Cuda, G., De Vivo, F., Di, B., Ciaramella,
F., Caputo, G., de Divitiis, O., 2000. Changes in myocardial cytoskeletal
intermediate filaments and myocyte contractile dysfunction in dilated
cardiomyopathy: an in vivo study in humans. Heart 84, 659–667.
Dong, Q., Chen, L., Lu, Q., Sharma, S., Li, L., Morimoto, S., Wang, G., 2014. Quercetin
attenuates doxorubicin cardiotoxicity by modulating Bmi-1 expression. Br. J.
Pharmacol. 171, 4440–4454.
Dudka, J., Gieroba, R., Korga, A., Burdan, F., Matysiak, W., Jodlowska-Jedrych, B.,
Mandziuk, S., Korobowicz, E., Murias, M., 2012. Different effects of resveratrol
on dose-related Doxorubicin-induced heart and liver toxicity. Evid. Based
Complement Alternat. Med. 2012, 606183.
El-Agamy, D.S., Abo-Haded, H.M., Elkablawy, M.A., 2016. Cardioprotective effects
of sitagliptin against doxorubicin-induced cardiotoxicity in rats. Exp. Biol. Med.
(Maywood) 241, 1577–1587.
Elbaky, N.A., Ali, A.A., Ahmed, R.A., 2010. Cardioprotective effect of simvastatin on
doxorubicininduced oxidative cardiotoxicity in rats. J. Basic Appl. Sci. 6, 29–38.
Elsherbiny, N.M., Salama, M.F., Said, E., El-Sherbiny, M., Al-Gayyar, M.M., 2016.
Crocin protects against doxorubicin-induced myocardial toxicity in rats
through down-regulation of inflammatory and apoptic pathways. Chem. Biol.
Interact. 247, 39–48.
Gala, A.A.A., 2013. Protective effect of Zingiber officinale (ginger) on doxorubicin
induced oxidative cardiotoxicity in rats. Life Sci. J. 10, 2924–2934.
Garrido-Maraver, J., Cordero, M.D., Oropesa-Ávila, M., Fernández Vega, A., De La
Mata, M., Delgado Pavón, A., De Miguel, M., Pérez Calero, C., Villanueva Paz, M.,
Cotán, D., 2014. Coenzyme q10 therapy. Mol. Syndromol. 5, 187–197.
Gianni, L., Salvatorelli, E., Minotti, G., 2007. Anthracycline cardiotoxicity in breast
cancer patients: synergism with trastuzumab and taxanes. Cardiovasc. Toxicol.
7, 67–71.
Goyal, S.N., Mahajan, U.B., Chandrayan, G., Kumawat, V.S., Kamble, S., Patil, P.,
Agrawal, Y.O., Patil, C.R., Ojha, S., 2016. Protective effect of oleanolic acid on
oxidative injury and cellular abnormalities in doxorubicin induced cardiac
toxicity in rats. Am. J. Transl. Res. 8, 60–69.
Guan, Y., Zheng, X., Yang, Z., Li, S., 2009. [Protective effects of L-carnitine upon
testicular ischemia-reperfusion damage in rats]. Zhonghua yi xue za zhi 89,
1858–1861.
Hadi, N., Yousif, N.G., Al-Amran, F.G., Huntei, N.K., Mohammad, B.I., Ali, S.J., 2012.
Vitamin E and telmisartan attenuates doxorubicin induced cardiac injury in rat
through down regulation of inflammatory response. BMC Cardiovasc. Disord.
12, 1.
Hajrasouliha, A.R., Tavakoli, S., Jabehdar-Maralani, P., Shafaroodi, H., Borhani, A.A.,
Houshmand, G., Sadeghipour, H., Dehghani, M., Dehpour, A.R., 2004. Resistance
of cholestatic rats against epinephrine-induced arrhythmia: the role of nitric
oxide and endogenous opioids. Eur. J. Pharmacol. 499, 307–313.
Heling, A., Zimmermann, R., Kostin, S., Maeno, Y., Hein, S., Devaux, B., Bauer, E.,
Klovekorn, W.P., Schlepper, M., Schaper, W., Schaper, J., 2000. Increased
expression of cytoskeletal, linkage, and extracellular proteins in failing human
myocardium. Circ. Res. 86, 846–853.
Ikeda, Y., Aihara, K., Akaike, M., Sato, T., Ishikawa, K., Ise, T., Yagi, S., Iwase, T., Ueda,
Y., Yoshida, S., Azuma, H., Walsh, K., Tamaki, T., Kato, S., Matsumoto, T., 2010.
Androgen receptor counteracts Doxorubicin-induced cardiotoxicity in male
mice. Mol. Endocrinol. 24, 1338–1348.
Jagetia, G., Venkatesh, P., 2015. An indigenous plant bael (Aegle marmelos (L.)
correa) extract protects against the doxorubicin-Induced cardiotoxicity in
mice. Biochem. Physiol. 4, 2.
Jones, S.P., Greer, J.J., Kakkar, A.K., Ware, P.D., Turnage, R.H., Hicks, M., van Haperen,
R., de Crom, R., Kawashima, S., Yokoyama, M., 2004. Endothelial nitric oxide
synthase overexpression attenuates myocardial reperfusion injury. Am. J.
Physiol.-Heart Circulatory Physiol. 286, H276–H282.
Kalyanaraman, B., Joseph, J., Kalivendi, S., Wang, S., Konorev, E., Kotamraju, S.,
2002. Doxorubicin-induced apoptosis: implications in cardiotoxicity. Mol. Cell.
Biochem. 234–235, 119–124.
Kwong, L.K., Kamzalov, S., Rebrin, I., Bayne, A.C., Jana, C.K., Morris, P., Forster, M.J.,
Sohal, R.S., 2002. Effects of coenzyme Q(10) administration on its tissue
concentrations, mitochondrial oxidant generation, and oxidative stress in the
rat. Free Radic. Biol. Med. 33, 627–638.
Ma, Y., de Castro Brás, L.E., Toba, H., Iyer, R.P., Hall, M.E., Winniford, M.D., Lange,
R.A., Tyagi, S.C., Lindsey, M.L., 2014. Myofibroblasts and the extracellular
matrix network in post-myocardial infarction cardiac remodeling. Pflugers
Arch. 466, 1113–1127.
Mandziuk, S., Gieroba, R., Korga, A., Matysiak, W., Jodlowska-Jedrych, B., Burdan, F.,
Poleszak, E., Kowalczyk, M., Grzycka-Kowalczyk, L., Korobowicz, E., Jozefczyk,
A., Dudka, J., 2015. The differential effects of green tea on dose-dependent
doxorubicin toxicity. Food Nutr. Res. 59, 29754.
17. 426 H.N. Mustafa et al. / Tissue and Cell 49 (2017) 410–426
Mantawy, E.M., El-Bakly, W.M., Esmat, A., Badr, A.M., El-Demerdash, E., 2014.
Chrysin alleviates acute doxorubicin cardiotoxicity in rats via suppression of
oxidative stress, inflammation and apoptosis. Eur. J. Pharmacol. 728, 107–118.
Mescka, C.P., Rosa, A.P., Schirmbeck, G., da Rosa, T.H., Catarino, F., de Souza, L.O.,
Guerreiro, G., Sitta, A., Vargas, C.R., Dutra-Filho, C.S., 2016. L-carnitine prevents
oxidative stress in the brains of rats subjected to a chemically induced chronic
model of MSUD. Mol. Neurobiol. 53, 6007–6017.
Miranda, K.M., Espey, M.G., Yamada, K., Krishna, M., Ludwick, N., Kim, S.,
Jourd’heuil, D., Grisham, M.B., Feelisch, M., Fukuto, J.M., 2001. Unique oxidative
mechanisms for the reactive nitrogen oxide species, nitroxyl anion. J. Biol.
Chem. 276, 1720–1727.
Mitry, M.A., Edwards, J.G., 2016. Doxorubicin induced heart failure: phenotype and
molecular mechanisms. Int. J. Cardiol. Heart Vasc. 10, 17–24.
Moshage, H., Kok, B., Huizenga, J.R., Jansen, P.L., 1995. Nitrite and nitrate
determinations in plasma: a critical evaluation. Clin. Chem. 41, 892–896.
Mustafa, H.N., Hussein, A.M., 2016. Does allicin combined with vitamin B-complex
have superior potentials than alpha-tocopherol alone in ameliorating lead
acetate-induced Purkinje cell alterations in rats? An immunohistochemical
and ultrastructural study. Folia Morphol (Warsz) 75 (1), 76–86.
Mustafa, H.N., El Awdan, S.A., Hegazy, G.A., Jaleel, G.A.A., 2015. Prophylactic role of
coenzyme Q10 and Cynara scolymus L on doxorubicin-induced toxicity in rats:
biochemical and immunohistochemical study. Indian J. Pharmacol. 47,
649–656.
Mustafa, H.N., 2015. Potential alleviation of chlorella vulgaris and zingiber
officinale on lead-Induced testicular toxicity: an ultrastructural study. Folia
Biol. (Praha) 63, 269–278.
Mustafa, H.N., 2016. The role of curcumin in streptozotocin-induced hepatic
damage and the trans-differentiation of hepatic stellate cells. Tissue Cell 48,
81–88.
Nascimento, A.R., Machado, M.V., Gomes, F., Vieira, A.B.,
Gonc¸ alves-de-Albuquerque, C.F., Lessa, M.A., Bousquet, P., Tibiric¸ á, E., 2016.
Central sympathetic modulation reverses microvascular alterations in a rat
model of high-Fat diet-Induced metabolic syndrome. Microcirculation 23,
320–329.
Naugle, J.E., Olson, E.R., Zhang, X., Mase, S.E., Pilati, C.F., Maron, M.B., Folkesson,
H.G., Horne, W.I., Doane, K.J., Meszaros, J.G., 2006. Type VI collagen induces
cardiac myofibroblast differentiation: implications for postinfarction
remodeling. Am. J. Physiol.-Heart Circulatory Physiol. 290, H323–H330.
Neilan, T.G., Blake, S.L., Ichinose, F., Raher, M.J., Buys, E.S., Jassal, D.S., Furutani, E.,
Perez-Sanz, T.M., Graveline, A., Janssens, S.P., 2007. Disruption of nitric oxide
synthase 3 protects against the cardiac injury, dysfunction, and mortality
induced by doxorubicin. Circulation 116, 506–514.
Octavia, Y., Tocchetti, C.G., Gabrielson, K.L., Janssens, S., Crijns, H.J., Moens, A.L.,
2012. Doxorubicin-induced cardiomyopathy: from molecular mechanisms to
therapeutic strategies. J. Mol. Cell. Cardiol. 52, 1213–1225.
Oktem, G., Uysal, A., Oral, O., Sezer, E.D., Olukman, M., Erol, A., Akgur, S.A., Bilir, A.,
2012. Resveratrol attenuates doxorubicin-induced cellular damage by
modulating nitric oxide and apoptosis. Exp. Toxicol. Pathol. 64, 471–479.
Olukman, M., Can, C., Erol, A., Öktem, G., Oral, O., 2009. Reversal of
doxorubicin-induced vascular dysfunction by resveratrol in rat thoracic aorta:
is there a possible role of nitric oxide synthase inhibition? Anatol. J. Cardiol. 9,
260–266.
Potemski, P., Polakowski P. a. Wiktorowska-Owczarek, A.K., Owczarek, J.,
Pluzanska, A., Orszulak-Michalak, D., 2006. Amifostine improves hemodynamic
parameters in doxorubicin-pretreated rabbits. Pharmacol. Rep. 58, 966.
Pott, C., Steinritz, D., Bolck, B., Mehlhorn, U., Brixius, K., Schwinger, R.H., Bloch, W.,
2006. eNOS translocation but not eNOS phosphorylation is dependent on
intracellular Ca2+ in human atrial myocardium. Am. J. Physiol. Cell Physiol.
290, C1437–1445.
Pradegan, N., Vida, V.L., Geva, T., Stellin, G., White, M.T., Sanders, S.P., Padera, R.F.,
2016. Myocardial histopathology in late-repaired and unrepaired adults with
tetralogy of Fallot. Cardiovasc. Pathol. 25, 225–231.
Rashikh, A., Abul Kalam, N., Akhtar, M., Mahmood, D., Pillai, K.K., Ahmad, S.J., 2011.
Protective effects of aliskiren in doxorubicin-induced acute cardiomyopathy in
rats. Hum. Exp. Toxicol. 30, 102–109.
Rastogi, S., Mishra, S., Zaca, V., Mika, Y., Rousso, B., Sabbah, H.N., 2008. Effects of
chronic therapy with cardiac contractility modulation electrical signals on
cytoskeletal proteins and matrix metalloproteinases in dogs with heart failure.
Cardiology 110, 230–237.
Reagan, W.J., York, M., Berridge, B., Schultze, E., Walker, D., Pettit, S., 2013.
Comparison of cardiac troponin I and T, including the evaluation of an
ultrasensitive assay, as indicators of doxorubicin-induced cardiotoxicity.
Toxicol. Pathol. 41, 1146–1158.
Sahu, B.D., Kumar, J.M., Kuncha, M., Borkar, R.M., Srinivas, R., Sistla, R., 2016.
Baicalein alleviates doxorubicin-induced cardiotoxicity via suppression of
myocardial oxidative stress and apoptosis in mice. Life Sci. 144, 8–18.
Schaper, J., Froede, R., Hein, S., Buck, A., Hashizume, H., Speiser, B., Friedl, A., Bleese,
N., 1991. Impairment of the myocardial ultrastructure and changes of the
cytoskeleton in dilated cardiomyopathy. Circulation 83, 504–514.
Sharov, V.G., Kostin, S., Todor, A., Schaper, J., Sabbah, H.N., 2005. Expression of
cytoskeletal, linkage and extracellular proteins in failing dog myocardium.
Heart Fail. Rev. 10, 297–303.
Sun, Z., Yan, B., Yu, W.Y., Yao, X., Ma, X., Sheng, G., Ma, Q., 2016. Vitexin attenuates
acute doxorubicin cardiotoxicity in rats via the suppression of oxidative stress,
inflammation and apoptosis and the activation of FOXO3a. Exp. Ther. Med. 12,
1879–1884.
Surai, P.F., 2015. Antioxidant action of carnitine: molecular mechanisms and
practical applications. EC Veterinary Sci. 2, 66–84.
Viswanatha Swamy, A.H., Wangikar, U., Koti, B.C., Thippeswamy, A.H., Ronad, P.M.,
Manjula, D.V., 2011. Cardioprotective effect of ascorbic acid on
doxorubicin-induced myocardial toxicity in rats. Indian J. Pharmacol. 43,
507–511.
Weber, K.T., Sun, Y., Bhattacharya, S.K., Ahokas, R.A., Gerling, I.C., 2013.
Myofibroblast-mediated mechanisms of pathological remodelling of the heart.
Nat. Rev. Cardiol. 10, 15–26.
Yin, W., Zhang, P., Huang, J.H., Zhang, Q.Y., Fan, R., Li, J., Zhou, J.J., Hu, Y.Z., Guo, H.T.,
Zhang, S.M., Wang, Y.M., Kaye, A.D., Gu, C.H., Liu, J.C., Cheng, L., Cui, Q., Yi, D.H.,
Pei, J.M., 2009. Stimulation of kappa-opioid receptor reduces
isoprenaline-induced cardiac hypertrophy and fibrosis. Eur. J. Pharmacol. 607,
135–142.
Zhang, S., Liu, X., Bawa-Khalfe, T., Lu, L.S., Lyu, Y.L., Liu, L.F., Yeh, E.T., 2012.
Identification of the molecular basis of doxorubicin-induced cardiotoxicity.
Nat. Med. 18, 1639–1642.
Zhang, Y.P., Song, C.Y., Yuan, Y., Eber, A., Rodriguez, Y., Levitt, R.C., Takacs, P., Yang,
Z., Goldberg, R., Candiotti, K.A., 2013. Diabetic neuropathic pain development
in type 2 diabetic mouse model and the prophylactic and therapeutic effects of
coenzyme Q10. Neurobiol. Dis. 58, 169–178.
de Salvi Guimaraes, F., de Moraes, W.M., Bozi, L.H., Souza, P.R., Antonio, E.L.,
Bocalini, D.S., Tucci, P.J., Ribeiro, D.A., Brum, P.C., Medeiros, A., 2017.
Dexamethasone-induced cardiac deterioration is associated with both calcium
handling abnormalities and calcineurin signaling pathway activation. Mol. Cell.
Biochem. 424, 87–98.