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Citation: Fatmi N, Fatima N, Shahzada MZ, Sharma S, Kumar R, Ali Mm, et al. Ameliorative Effect of Murraya
Koenigii on Arsenic Induced Toxicity in Swiss Albino Mice. Austin J Pharmacol Ther. 2017; 5(3).1097.
Austin J Pharmacol Ther - Volume 5 Issue 3 - 2017
ISSN: 2373-6208 | www.austinpublishinggroup.com
Kumar et al. © All rights are reserved
Austin Journal of Pharmacology and
Therapeutics
Open Access
Abstract
Arsenic is a metalloid that can be toxic to humans and other living organisms,
occurs naturally and anthropogenically throughout the world at varying
concentrations, including concentrations of concern in soil or groundwater.
Presently, in Bihar (India) 18 districts are affected with arsenic poisoning in
ground water causing lots of health hazards among the population. This arsenic
intoxication has caused lots of health related problems in the population. The
present study has been designed, to study the ameliorative effect of Murraya
Koenigii on sodium arsenite induced toxicity in Swiss albino mice. The treatment
groups received sodium arseniteorally at the dose of 3mgkg-1 body weight daily
for 4 weeks followed by administration of Murraya Koenigii 350mgkg-1 body
weight daily by gavage method for 4& 6 weeks. Their biochemical levels like
liver and kidney function tests were assayed and were observed with elevated
levels. Furthermore, their free radical assessment like lipid peroxidation levels
were assayed which was found to be many folds higher. But, after administration
of aqueous extract of Murraya Koenigii, there was significant amelioration in
the biochemical and lipid peroxidation levels. Therefore, it is evident from
the present study that Murraya Koenigii possesses antidote effects and acts
effectively against arsenic induced toxicity.
Keywords: Arsenic; Toxicity; Antidote effect; Swiss albino mice
In Sidhha system of medicine it’s known as Karuveppilai and
used as herb in Ayurvedic medicine. The leaves of Murraya Koenigii
are also used as herb in Ayurvedic medicine. Their properties include
much value as an antidiabetic, antioxidant, antimicrobial, anti-
inflammatory hepatoprotective, anti- hyper cholesterol lemic, and etc
[10,11]. Curry leaves are also known to be good for hair, for keeping
it healthy and long as it also contains iron. Although, most commonly
used in curries leaves from the curry tree can be used in many other
dishes to add spice. The essential oil from leaves of Murraya Koenigii
exhibit strong antibacterial as well as antifungal activity [12-14].
Present study aims to illustrate the ameliorative effect Murraya
Koenigii on arsenic induced toxicity in mice.
Materials and Methods
Animals
Swiss albino mice (Mus musculus), weighing 30g to 35g of 8
weeks old, were obtained from animal house of Mahavir Cancer
Institute and Research Centre, Patna, India (CPCSEA Regd-No.
1129/bc/07/CPCSEA). The research work was approved by the IAEC
(Institutional Animal Ethics Committee) with IAEC No. 2015/3D-
16/12/15. Food and water to mice were provided ad libitum (prepared
mixed formulated food by the laboratory itself). The experimental
animals were housed in conventional polypropylene cages in small
groups (2 each). The mice were randomly assigned to control and
treatment groups. The temperature in the experimental animal room
was maintained at 22±2θ
C with 12h light/dark cycle.
Chemicals
Sodium Arsenite (98.5%) manufactured by Sigma-Aldrich, USA
Introduction
Arsenic is a metalloid that can be toxic to humans and other living
organisms, occurs naturally and anthropogenically throughout the
world at varying concentrations, including concentrations of concern
in soil or groundwater. Acute arsenic exposure harms human health
in many ways including the development of malignancies, severe
gastrointestinal toxicities, diabetes, cardiac arrhythmias, cancer
and death [1]. Natural sources such as volcanic action, erosion of
rocks, and forest fires introduce arsenic into the environment (EPA,
2001). Anthropogenic sources include arsenic added to the soil plant
system as insecticides, herbicides, pesticides, livestock dips and wood
preservatives. It is estimated that more than 200 million people
worldwide are chronically exposed to dangerous levels of arsenic
which leads to several diseases, including various types of cancer
[2]. Arsenical exposure through drinking water is common in many
areas of the world [3]. Metabolic disorders, hypertrophy of adrenal
glands [4] and anaemia [5], inhibition of the activity of steroidogenic
enzymes [6] and reduction in the weight of the testis and accessory
sex organs [7] are associated with exposure to arsenicals. Presently,
in Bihar, 18 districts are highly affected with arsenic poisoning. The
groundwater contamination by arsenic is maximum 1928ppb in
Buxar district of Bihar [8,9].
Traditional medicines include herbal medicines composed of
herbs, herbal materials, herbal preparations, and finished herbal
products, that contain as active ingredients parts of plants, or other
plant materials, or combinations thereof. Since last two decades, the
phytoremediation of various heavy metals borne diseases has gained
special attention to researchers.
Fatmi N1
, Fatima N1
, Shahzada MZ1
, Sharma S1
,
Kumar R2
, Ali M2
and Kumar A2
*
1
Department of Biochemistry, Magadh University, Bodh
Gaya, Bihar, India
2
Mahavir Cancer Institute & Research Centre, Phulwari
Sharif, Patna, India
*Corresponding author: Arun Kumar, Mahavir
Cancer Institute & Research Centre, Phulwari Sharif,
Patna, India
Received: August 30, 2017; Accepted: October 05,
2017; Published: October 12, 2017
Research Article
Ameliorative Effect of Murraya Koenigii on Arsenic
Induced Toxicity in Swiss Albino Mice
Austin J Pharmacol Ther 5(3): id1097 (2017) - Page - 02
Kumar A Austin Publishing Group
Submit your Manuscript | www.austinpublishinggroup.com
(CAS Number: 7784-46-5), was obtained from the Scientific store of
Patna of Bihar India.
Preparation of Murraya Koenigii aqueous extract: In the present
study, leaves of Murraya Koenigii were procured locally from Patna,
Bihar, India The identity of the medicinal plant was confirmed by Dr.
Ramakant Pandey (Botanist), Department of Biochemistry, Patna
University, Patna, Bihar, India. The collected leaves of Murraya
Koenigii were shade dried and were grinded to fine powder. The
aqueous extract dose was calculated after LD50 estimation which was
found to be 4800mgkg-1
body weight and the final dose was fixed to
350mgkg-1
body weight.
Study groups & sampling
The control group of 6 mice received distilled water as drinking
water. The treatment groups (n= 18) received Sodium arsenite daily
at the dose of 3mgkg-1
body weight for 4 weeks orally (after estimation
of LD50 value which was found to be 8mgkg-1
body weight) followed
by administration of Murraya Koenigii 350mgkg-1
body weight
daily by gavage method for 4and 6 weeks. Mice were sacrificed after
completion of their treatment and their blood were collected and
serum were extracted for biochemical assays and lipid peroxidation
estimation.
Biochemical evaluation
The Liver Function Test (LFT) were assayed by methods as
Serum Glutamate Pyruvate Transaminase (SGPT), Serum Glutamate
Oxalate Transaminase (SGOT) [15], Alkaline Phosphatase ALP [16],
Bilirubin [17].The Kidney Function Test (KFT) were assayed by
methods as Urea by [18,19] Uric acid by [20] and Creatinine by [21].
Lipid Peroxidation (LPO)
Thiobarbituric acid reactive substances (TBARS), as a marker
for LPO, were determined by the double heating method [22]. The
principle of the method was a spectrophotometric measurement of
the color produced during the reaction to thiobarbituric acid (TBA)
with malondialdehyde (MDA). For this purpose, 2.5ml of 100g/l
trichloroacetic acid (TCA) solution was added to 0.5ml serum in a
centrifuge tube and incubated for 15min at 90θ
C. After cooling in tap
water, the mixture was centrifuged at 3000g for 10min, and 2ml of
the supernatant was added to 1ml of 6.7g/l TBA solution in a test tube
and again incubated for 15min at 90θ
C. The solution was then cooled
in tap water and its absorbance was measured using Thermo Scientific
UV-10 (UV –Vis) spectrophotometer (USA) at 532nm.
Statistical analysis
Results are presented as mean ±SD and total variation present
in a set of data was analysed through one way analysis of variance
(ANOVA). Difference among mean values has been analysed by
applying Dunnet’s t-test. Calculations were performed with the
Graph Pad Prism Program (Graph Pad software, Inc., San Diego,
U.S.A.). The criterion for statistical significance was set at P< 0.05.
Results
The liver function test parameters - SGPT levels in control mice
was 25.67 ± 1.20 U/ml while in arsenic four weeks treated group
it was 150.3 ± 2.33 U/ml (Graph Figure 1). In Murraya Koenigii
administered group of mice SGPT levels was 124.33 ± 2.33 U/ml after
4 weeks and 47.33 ± 1.90 U/ml after 6 weeks. SGOT levels in control
mice was 22.00 ± 2.64 U/ml while in arsenic four weeks treated group
it was 128.7 ± 2.60 U/ml (Graph Figure 2). In Murraya Koenigii
administered group of mice SGOT levels was 83.67 ± 2.40 U/ml after
4 weeks and 33.67 ± 2.87 U/ml after 6 weeks. ALP levels in control
mice was 6.567 ± 0.35 K.A units, while in arsenic four weeks treated
Graph Figure 1: Effect of Murraya Koenigii on Arsenic induced toxicity
showing SGPT activity (n=6, values are mean± S.D).
Graph Figure 2: Effect of Murraya Koenigii on Arsenic induced toxicity
showing SGOT activity (n=6, values are mean± S.D).
Graph Figure 3: Effect of Murraya Koenigii on Arsenic induced toxicity
showing ALP activity (n=6, values are mean± S.D).
Austin J Pharmacol Ther 5(3): id1097 (2017) - Page - 03
Kumar A Austin Publishing Group
Submit your Manuscript | www.austinpublishinggroup.com
group it was 29.67 ± 1.76 K.A units (Graph Figure 3). In Murraya
Koenigii administered group of mice ALP levels was 18.77 ± 0.52 K.A.
units after 4 weeks and 11.77 ± 0.57 K.A. units after 6 weeks. Bilirubin
levels in control mice was 0.5633 ± 0.029 mg/dl while in arsenic four
weeks treated group it was 1.677 ± 0.06 mg/dl (Graph Figure 4). In
Murraya Koenigii administered group of mice bilirubin levels was
1.2133 ± 0.02 mg/dl after 4 weeks and 0.9133 ± 0.08 mg/dl after 6
weeks.
The Kidney Function Tests - Urea levels in control mice was 17.33
± 2.02 mg/dl while in arsenic four weeks treated group it was 86.67 ±
2.96 mg/dl (Graph Figure 5). In Murraya Koenigii administered group
of mice urea levels was 57.16 ± 1.13 mg/dl after 4 weeks and 35.67 ±
1.24 mg/dl after 6 weeks. Uric acid levels in control mice was 4.800 ±
0.45 mg/dl while in arsenic our weeks treated group it was 9.733 ± 0.23
mg/dl (Graph Figure 6). In Murraya Koenigii administered group of
mice uric acid levels was 8.201 ± 0.28 mg/dl after 4 weeks and 7.033 ±
0.17 mg/dl after 6 weeks. Creatinine levels in control mice was 0.5167
± 0.03 mg% while in arsenic four weeks treated group it was 3.733 ±
0.23 mg% (Graph Figure 7 in Murraya Koenigii administered group
of mice creatinine levels was 2.856 ± 0.17 mg% and after 4 weeks and
1.972 ± 0.24 mg% after 6 weeks.
Lipid peroxidation levels in control mice was 1.987 ± 0.06 nmol/
ml while in arsenic four weeks treated group it was 58.73 ± 1.36 nmol/
ml (Graph Figure 8). In Murraya Koenigii administered group of
mice Lipid peroxidation levels was 38.27 ± 0.32 nmol/ml after 4 weeks
and 14.43 ± 0.40 nmol/ml after 6 weeks.
Graph Figure 4: Effect of Murraya Koenigii on Arsenic induced toxicity
showing Bilirubin activity (n=6, values are mean± S.D).
Graph Figure 5: Effect of Murraya Koenigii on Arsenic induced toxicity
showing Urea activity (n=6, values are mean± S.D).
Graph Figure 6: Effect of Murraya Koenigii on Arsenic induced toxicity
showing Uric Acid activity (n=6, values are mean± S.D).
Graph Figure 7: Effect of Murraya Koenigii on Arsenic induced toxicity
showing Creatinine activity (n=6, values are mean± S.D).
Graph Figure 8: Effect of Murraya Koenigii on Arsenic induced toxicity
showing Lipid peroxidation activity (n=6, values are mean± S.D).
Austin J Pharmacol Ther 5(3): id1097 (2017) - Page - 04
Kumar A Austin Publishing Group
Submit your Manuscript | www.austinpublishinggroup.com
Discussion
Liver is one of the important organ actively involved in metabolic
functions and is in frequent target of number of toxicants. The main
function of the liver is detoxification of toxins and xenobiotics [23].
Because liver performs many vital functions in the human body,
damage of liver causes serious problems [24]. SGPT and SGOT are
the most often used and most specific indicators of hepatic injury
and represent markers of hepatocellular disease [25]. Arsenic is
known to produce damages in liver function [26]. SGPT and SGOT
are reliable determinants of liver parenchymal injury [27] and their
activities are significantly increased in arsenic treated mice indicating
liver dysfunction. Assay of serum ALP activity has been recognized
as a suitable marker of skeletal and hepatobiliary disorder. Moreover,
an elevated serum level of ALP activity is frequently associated with
various pathological conditions [28,29]. Alkaline phosphate is a
non-specific tissue enzyme widely spread, mainly in the osteoblasts,
liver and biliary canaliculi [30,31]. Bilirubin is the conventional
indicator of liver diseases [32]. Hyperbilirubinemia is a very sensitive
test to substantiate the functional integrity of the liver and severity
of necrosis which increases the binding, conjugating and excretory
capacity of hepatocytes that is proportional to the erythrocyte
degeneration rate [33].
In the present study, administration of Sodium arsenite treated
mice showed insignificant increase in the levels of SGPT, SGOT,
ALP and bilirubin activities when compared with control mice. Oral
administration of aqueous extract of Murraya Koenigii (350 mg/kg-1
body weight) to sodium arsenite treated mice showed an inhibition
in the elevated levels of serum SGPT, SGOT, ALP and Bilirubin than
sodium arsenite alone treated mice.
The protective nature of M. Koenigii leaves extract was well
studied [34,35]. The effect attributed to the combined effect of
carbazole alkaloids – Mahanimbine, Girinimbine, Isomahanimbine,
murrayazoline, Murrayazolidine, Mahanine and ascorbic acid,
α-tocopherol and mineral (Zn, Cu, Fe) contents of M.Koenigii
leaves extract. This study proved M. Koenigii a promising and a rich
source of free radical quenchers, which have been mediated through
hepatocyte membrane stabilizing activity along with the reduction of
fat metabolism [36]. The normal morphology of cell was maintained
after ethanolic challenge when aqueous extract containing tannins
and carbazole alkaloids of M. Koenigii was given.
In kidney, arsenic exerts its deleterious toxic effects through
several mechanisms, the most significant of which is the reversible
combination with suflydryl group of proteins present in glomerular
filtration membrane [37]. In the present study also, there was decrease
in the levels of kidney function tests especially the urea, uric acid and
creatinine levels denotes the ameliorative effect. Aqueous extract
of leaves produced a significant dose-dependent decrease in serum
urea and creatinine levels [38]. Arsenic causes oxidative damage by
producing reactive oxygen species [39,40] which damage proteins.
Due to lipophilic in nature, arsenic also attaches to lipid, increases
the lipid peroxidation [41] resulting in deposition of lipid droplets
in the slit pores of glomerular filtration membrane. Both the reasons
discussed above cause decreased Glomerular Filtration Rate (GFR),
thereby causing accumulation of nitrogenous waste product in blood.
In the present study also, there was immense increase in the levels of
lipid peroxidation (MDA) levels denotes the similar effect but there
was declination in the levels of the same by the ameliorative effect of
aqueous extract of Murraya Koenigii denotes the antioxidant activity.
The entire study therefore denotes the ameliorative effect of Murraya
Koenigii [42-45].
Conclusion
It is evident from study that Murraya Koenigii acts effectively
againstarsenicinducedtoxicity.Itrestorestheliverfunctiontestaswell
as kidney function test effectively. It also restores lipid peroxidation
levels up to the normal levels, thus indicates that Murraya Koenigii
possesses ameliorative effect against arsenic induced toxicity.
Acknowledgement
The authors are thankful to Mahavir Cancer Institute and
Research Centre for providing infrastructural facility during this
work.
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Citation: Fatmi N, Fatima N, Shahzada MZ, Sharma S, Kumar R, Ali Mm, et al. Ameliorative Effect of Murraya
Koenigii on Arsenic Induced Toxicity in Swiss Albino Mice. Austin J Pharmacol Ther. 2017; 5(3).1097.
Austin J Pharmacol Ther - Volume 5 Issue 3 - 2017
ISSN: 2373-6208 | www.austinpublishinggroup.com
Kumar et al. © All rights are reserved

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Murraya Koenigii Reduces Arsenic Toxicity in Mice

  • 1. Citation: Fatmi N, Fatima N, Shahzada MZ, Sharma S, Kumar R, Ali Mm, et al. Ameliorative Effect of Murraya Koenigii on Arsenic Induced Toxicity in Swiss Albino Mice. Austin J Pharmacol Ther. 2017; 5(3).1097. Austin J Pharmacol Ther - Volume 5 Issue 3 - 2017 ISSN: 2373-6208 | www.austinpublishinggroup.com Kumar et al. © All rights are reserved Austin Journal of Pharmacology and Therapeutics Open Access Abstract Arsenic is a metalloid that can be toxic to humans and other living organisms, occurs naturally and anthropogenically throughout the world at varying concentrations, including concentrations of concern in soil or groundwater. Presently, in Bihar (India) 18 districts are affected with arsenic poisoning in ground water causing lots of health hazards among the population. This arsenic intoxication has caused lots of health related problems in the population. The present study has been designed, to study the ameliorative effect of Murraya Koenigii on sodium arsenite induced toxicity in Swiss albino mice. The treatment groups received sodium arseniteorally at the dose of 3mgkg-1 body weight daily for 4 weeks followed by administration of Murraya Koenigii 350mgkg-1 body weight daily by gavage method for 4& 6 weeks. Their biochemical levels like liver and kidney function tests were assayed and were observed with elevated levels. Furthermore, their free radical assessment like lipid peroxidation levels were assayed which was found to be many folds higher. But, after administration of aqueous extract of Murraya Koenigii, there was significant amelioration in the biochemical and lipid peroxidation levels. Therefore, it is evident from the present study that Murraya Koenigii possesses antidote effects and acts effectively against arsenic induced toxicity. Keywords: Arsenic; Toxicity; Antidote effect; Swiss albino mice In Sidhha system of medicine it’s known as Karuveppilai and used as herb in Ayurvedic medicine. The leaves of Murraya Koenigii are also used as herb in Ayurvedic medicine. Their properties include much value as an antidiabetic, antioxidant, antimicrobial, anti- inflammatory hepatoprotective, anti- hyper cholesterol lemic, and etc [10,11]. Curry leaves are also known to be good for hair, for keeping it healthy and long as it also contains iron. Although, most commonly used in curries leaves from the curry tree can be used in many other dishes to add spice. The essential oil from leaves of Murraya Koenigii exhibit strong antibacterial as well as antifungal activity [12-14]. Present study aims to illustrate the ameliorative effect Murraya Koenigii on arsenic induced toxicity in mice. Materials and Methods Animals Swiss albino mice (Mus musculus), weighing 30g to 35g of 8 weeks old, were obtained from animal house of Mahavir Cancer Institute and Research Centre, Patna, India (CPCSEA Regd-No. 1129/bc/07/CPCSEA). The research work was approved by the IAEC (Institutional Animal Ethics Committee) with IAEC No. 2015/3D- 16/12/15. Food and water to mice were provided ad libitum (prepared mixed formulated food by the laboratory itself). The experimental animals were housed in conventional polypropylene cages in small groups (2 each). The mice were randomly assigned to control and treatment groups. The temperature in the experimental animal room was maintained at 22±2θ C with 12h light/dark cycle. Chemicals Sodium Arsenite (98.5%) manufactured by Sigma-Aldrich, USA Introduction Arsenic is a metalloid that can be toxic to humans and other living organisms, occurs naturally and anthropogenically throughout the world at varying concentrations, including concentrations of concern in soil or groundwater. Acute arsenic exposure harms human health in many ways including the development of malignancies, severe gastrointestinal toxicities, diabetes, cardiac arrhythmias, cancer and death [1]. Natural sources such as volcanic action, erosion of rocks, and forest fires introduce arsenic into the environment (EPA, 2001). Anthropogenic sources include arsenic added to the soil plant system as insecticides, herbicides, pesticides, livestock dips and wood preservatives. It is estimated that more than 200 million people worldwide are chronically exposed to dangerous levels of arsenic which leads to several diseases, including various types of cancer [2]. Arsenical exposure through drinking water is common in many areas of the world [3]. Metabolic disorders, hypertrophy of adrenal glands [4] and anaemia [5], inhibition of the activity of steroidogenic enzymes [6] and reduction in the weight of the testis and accessory sex organs [7] are associated with exposure to arsenicals. Presently, in Bihar, 18 districts are highly affected with arsenic poisoning. The groundwater contamination by arsenic is maximum 1928ppb in Buxar district of Bihar [8,9]. Traditional medicines include herbal medicines composed of herbs, herbal materials, herbal preparations, and finished herbal products, that contain as active ingredients parts of plants, or other plant materials, or combinations thereof. Since last two decades, the phytoremediation of various heavy metals borne diseases has gained special attention to researchers. Fatmi N1 , Fatima N1 , Shahzada MZ1 , Sharma S1 , Kumar R2 , Ali M2 and Kumar A2 * 1 Department of Biochemistry, Magadh University, Bodh Gaya, Bihar, India 2 Mahavir Cancer Institute & Research Centre, Phulwari Sharif, Patna, India *Corresponding author: Arun Kumar, Mahavir Cancer Institute & Research Centre, Phulwari Sharif, Patna, India Received: August 30, 2017; Accepted: October 05, 2017; Published: October 12, 2017 Research Article Ameliorative Effect of Murraya Koenigii on Arsenic Induced Toxicity in Swiss Albino Mice
  • 2. Austin J Pharmacol Ther 5(3): id1097 (2017) - Page - 02 Kumar A Austin Publishing Group Submit your Manuscript | www.austinpublishinggroup.com (CAS Number: 7784-46-5), was obtained from the Scientific store of Patna of Bihar India. Preparation of Murraya Koenigii aqueous extract: In the present study, leaves of Murraya Koenigii were procured locally from Patna, Bihar, India The identity of the medicinal plant was confirmed by Dr. Ramakant Pandey (Botanist), Department of Biochemistry, Patna University, Patna, Bihar, India. The collected leaves of Murraya Koenigii were shade dried and were grinded to fine powder. The aqueous extract dose was calculated after LD50 estimation which was found to be 4800mgkg-1 body weight and the final dose was fixed to 350mgkg-1 body weight. Study groups & sampling The control group of 6 mice received distilled water as drinking water. The treatment groups (n= 18) received Sodium arsenite daily at the dose of 3mgkg-1 body weight for 4 weeks orally (after estimation of LD50 value which was found to be 8mgkg-1 body weight) followed by administration of Murraya Koenigii 350mgkg-1 body weight daily by gavage method for 4and 6 weeks. Mice were sacrificed after completion of their treatment and their blood were collected and serum were extracted for biochemical assays and lipid peroxidation estimation. Biochemical evaluation The Liver Function Test (LFT) were assayed by methods as Serum Glutamate Pyruvate Transaminase (SGPT), Serum Glutamate Oxalate Transaminase (SGOT) [15], Alkaline Phosphatase ALP [16], Bilirubin [17].The Kidney Function Test (KFT) were assayed by methods as Urea by [18,19] Uric acid by [20] and Creatinine by [21]. Lipid Peroxidation (LPO) Thiobarbituric acid reactive substances (TBARS), as a marker for LPO, were determined by the double heating method [22]. The principle of the method was a spectrophotometric measurement of the color produced during the reaction to thiobarbituric acid (TBA) with malondialdehyde (MDA). For this purpose, 2.5ml of 100g/l trichloroacetic acid (TCA) solution was added to 0.5ml serum in a centrifuge tube and incubated for 15min at 90θ C. After cooling in tap water, the mixture was centrifuged at 3000g for 10min, and 2ml of the supernatant was added to 1ml of 6.7g/l TBA solution in a test tube and again incubated for 15min at 90θ C. The solution was then cooled in tap water and its absorbance was measured using Thermo Scientific UV-10 (UV –Vis) spectrophotometer (USA) at 532nm. Statistical analysis Results are presented as mean ±SD and total variation present in a set of data was analysed through one way analysis of variance (ANOVA). Difference among mean values has been analysed by applying Dunnet’s t-test. Calculations were performed with the Graph Pad Prism Program (Graph Pad software, Inc., San Diego, U.S.A.). The criterion for statistical significance was set at P< 0.05. Results The liver function test parameters - SGPT levels in control mice was 25.67 ± 1.20 U/ml while in arsenic four weeks treated group it was 150.3 ± 2.33 U/ml (Graph Figure 1). In Murraya Koenigii administered group of mice SGPT levels was 124.33 ± 2.33 U/ml after 4 weeks and 47.33 ± 1.90 U/ml after 6 weeks. SGOT levels in control mice was 22.00 ± 2.64 U/ml while in arsenic four weeks treated group it was 128.7 ± 2.60 U/ml (Graph Figure 2). In Murraya Koenigii administered group of mice SGOT levels was 83.67 ± 2.40 U/ml after 4 weeks and 33.67 ± 2.87 U/ml after 6 weeks. ALP levels in control mice was 6.567 ± 0.35 K.A units, while in arsenic four weeks treated Graph Figure 1: Effect of Murraya Koenigii on Arsenic induced toxicity showing SGPT activity (n=6, values are mean± S.D). Graph Figure 2: Effect of Murraya Koenigii on Arsenic induced toxicity showing SGOT activity (n=6, values are mean± S.D). Graph Figure 3: Effect of Murraya Koenigii on Arsenic induced toxicity showing ALP activity (n=6, values are mean± S.D).
  • 3. Austin J Pharmacol Ther 5(3): id1097 (2017) - Page - 03 Kumar A Austin Publishing Group Submit your Manuscript | www.austinpublishinggroup.com group it was 29.67 ± 1.76 K.A units (Graph Figure 3). In Murraya Koenigii administered group of mice ALP levels was 18.77 ± 0.52 K.A. units after 4 weeks and 11.77 ± 0.57 K.A. units after 6 weeks. Bilirubin levels in control mice was 0.5633 ± 0.029 mg/dl while in arsenic four weeks treated group it was 1.677 ± 0.06 mg/dl (Graph Figure 4). In Murraya Koenigii administered group of mice bilirubin levels was 1.2133 ± 0.02 mg/dl after 4 weeks and 0.9133 ± 0.08 mg/dl after 6 weeks. The Kidney Function Tests - Urea levels in control mice was 17.33 ± 2.02 mg/dl while in arsenic four weeks treated group it was 86.67 ± 2.96 mg/dl (Graph Figure 5). In Murraya Koenigii administered group of mice urea levels was 57.16 ± 1.13 mg/dl after 4 weeks and 35.67 ± 1.24 mg/dl after 6 weeks. Uric acid levels in control mice was 4.800 ± 0.45 mg/dl while in arsenic our weeks treated group it was 9.733 ± 0.23 mg/dl (Graph Figure 6). In Murraya Koenigii administered group of mice uric acid levels was 8.201 ± 0.28 mg/dl after 4 weeks and 7.033 ± 0.17 mg/dl after 6 weeks. Creatinine levels in control mice was 0.5167 ± 0.03 mg% while in arsenic four weeks treated group it was 3.733 ± 0.23 mg% (Graph Figure 7 in Murraya Koenigii administered group of mice creatinine levels was 2.856 ± 0.17 mg% and after 4 weeks and 1.972 ± 0.24 mg% after 6 weeks. Lipid peroxidation levels in control mice was 1.987 ± 0.06 nmol/ ml while in arsenic four weeks treated group it was 58.73 ± 1.36 nmol/ ml (Graph Figure 8). In Murraya Koenigii administered group of mice Lipid peroxidation levels was 38.27 ± 0.32 nmol/ml after 4 weeks and 14.43 ± 0.40 nmol/ml after 6 weeks. Graph Figure 4: Effect of Murraya Koenigii on Arsenic induced toxicity showing Bilirubin activity (n=6, values are mean± S.D). Graph Figure 5: Effect of Murraya Koenigii on Arsenic induced toxicity showing Urea activity (n=6, values are mean± S.D). Graph Figure 6: Effect of Murraya Koenigii on Arsenic induced toxicity showing Uric Acid activity (n=6, values are mean± S.D). Graph Figure 7: Effect of Murraya Koenigii on Arsenic induced toxicity showing Creatinine activity (n=6, values are mean± S.D). Graph Figure 8: Effect of Murraya Koenigii on Arsenic induced toxicity showing Lipid peroxidation activity (n=6, values are mean± S.D).
  • 4. Austin J Pharmacol Ther 5(3): id1097 (2017) - Page - 04 Kumar A Austin Publishing Group Submit your Manuscript | www.austinpublishinggroup.com Discussion Liver is one of the important organ actively involved in metabolic functions and is in frequent target of number of toxicants. The main function of the liver is detoxification of toxins and xenobiotics [23]. Because liver performs many vital functions in the human body, damage of liver causes serious problems [24]. SGPT and SGOT are the most often used and most specific indicators of hepatic injury and represent markers of hepatocellular disease [25]. Arsenic is known to produce damages in liver function [26]. SGPT and SGOT are reliable determinants of liver parenchymal injury [27] and their activities are significantly increased in arsenic treated mice indicating liver dysfunction. Assay of serum ALP activity has been recognized as a suitable marker of skeletal and hepatobiliary disorder. Moreover, an elevated serum level of ALP activity is frequently associated with various pathological conditions [28,29]. Alkaline phosphate is a non-specific tissue enzyme widely spread, mainly in the osteoblasts, liver and biliary canaliculi [30,31]. Bilirubin is the conventional indicator of liver diseases [32]. Hyperbilirubinemia is a very sensitive test to substantiate the functional integrity of the liver and severity of necrosis which increases the binding, conjugating and excretory capacity of hepatocytes that is proportional to the erythrocyte degeneration rate [33]. In the present study, administration of Sodium arsenite treated mice showed insignificant increase in the levels of SGPT, SGOT, ALP and bilirubin activities when compared with control mice. Oral administration of aqueous extract of Murraya Koenigii (350 mg/kg-1 body weight) to sodium arsenite treated mice showed an inhibition in the elevated levels of serum SGPT, SGOT, ALP and Bilirubin than sodium arsenite alone treated mice. The protective nature of M. Koenigii leaves extract was well studied [34,35]. The effect attributed to the combined effect of carbazole alkaloids – Mahanimbine, Girinimbine, Isomahanimbine, murrayazoline, Murrayazolidine, Mahanine and ascorbic acid, α-tocopherol and mineral (Zn, Cu, Fe) contents of M.Koenigii leaves extract. This study proved M. Koenigii a promising and a rich source of free radical quenchers, which have been mediated through hepatocyte membrane stabilizing activity along with the reduction of fat metabolism [36]. The normal morphology of cell was maintained after ethanolic challenge when aqueous extract containing tannins and carbazole alkaloids of M. Koenigii was given. In kidney, arsenic exerts its deleterious toxic effects through several mechanisms, the most significant of which is the reversible combination with suflydryl group of proteins present in glomerular filtration membrane [37]. In the present study also, there was decrease in the levels of kidney function tests especially the urea, uric acid and creatinine levels denotes the ameliorative effect. Aqueous extract of leaves produced a significant dose-dependent decrease in serum urea and creatinine levels [38]. Arsenic causes oxidative damage by producing reactive oxygen species [39,40] which damage proteins. Due to lipophilic in nature, arsenic also attaches to lipid, increases the lipid peroxidation [41] resulting in deposition of lipid droplets in the slit pores of glomerular filtration membrane. Both the reasons discussed above cause decreased Glomerular Filtration Rate (GFR), thereby causing accumulation of nitrogenous waste product in blood. In the present study also, there was immense increase in the levels of lipid peroxidation (MDA) levels denotes the similar effect but there was declination in the levels of the same by the ameliorative effect of aqueous extract of Murraya Koenigii denotes the antioxidant activity. The entire study therefore denotes the ameliorative effect of Murraya Koenigii [42-45]. Conclusion It is evident from study that Murraya Koenigii acts effectively againstarsenicinducedtoxicity.Itrestorestheliverfunctiontestaswell as kidney function test effectively. It also restores lipid peroxidation levels up to the normal levels, thus indicates that Murraya Koenigii possesses ameliorative effect against arsenic induced toxicity. Acknowledgement The authors are thankful to Mahavir Cancer Institute and Research Centre for providing infrastructural facility during this work. References 1. Platanias LC. Biological responses to arsenic compounds. Journal of Biological Chemistry. 2009; 284: 18583-18587. 2. 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Biomarkers of oxidative stress and heavy metal levels as indicators of environmental pollution in African cat fish (Clariasgariepinus) from Nigeria Ogun river. Int. J. Environ. Res. Pub. Hlth. 2007; 4: 158-165. 42. Environmental Protection Agency. Drinking water standards for Arsenic. United States Environmental Protection Agency 815-F-00-015. 2001. 43. Chakraborti D, Rahman MM, Ahamed S, Dutta RN, Pati S, Mukherjee SC. Arsenic groundwater contamination and its health effects in Patna district (capital of Bihar) in the middle Ganga plain, India. Chemosphere. 2016; 152: 520-529. 44. Chatterjee A, Das D, Chatter D. The study of underground water contamination by arsenic in the residential area of Behela and Calcutta, due to industrial pollution. Environmental Pollution. 1993; 80: 57-65. 45. Sarkar R, Mohanakumar KP, Chowdhury M. Effects of an organophosphate pesticide, quinalphos, on the hypothalamogonadal axis in adult male rats. J Reprod Fertil. 2000; 118: 29-38. Citation: Fatmi N, Fatima N, Shahzada MZ, Sharma S, Kumar R, Ali Mm, et al. Ameliorative Effect of Murraya Koenigii on Arsenic Induced Toxicity in Swiss Albino Mice. Austin J Pharmacol Ther. 2017; 5(3).1097. Austin J Pharmacol Ther - Volume 5 Issue 3 - 2017 ISSN: 2373-6208 | www.austinpublishinggroup.com Kumar et al. © All rights are reserved