Presentation by neha jain on aspirin induced alterations on liver and kidney
“Aspirin induced alterations in the Liver, Kidney and Reproductive System of female albino rat, Rattus rattus norvegicus”. 2010-2011 Supervisor Co-supervisor Presented ByProf. Arun Raghuwanshi Prof. Vinoy K. Shrivastava Neha Jain Faculty of Life Science Head of the M.Phil.Bioscience Department of Bioscience
CONTENTS INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION SUMMARY REFERANCES
INTRODUCTION • What is Aspirin • History of Aspirin • Mechanism of action • Common-side effects • Indications
What is AspirinAspirin is an analgesic the brand name Aspirin wascoined by the Bayer company of Germany also called acetylsalicylic acid most frequently selledacetylsalicylic acid compound Worldwide Aspirin belongs to a class of drugs called non-steroidal anti-inflammatory drugs (NSAIDs). They work by inhibiting the enzymes that make prostaglandins, chemicals that promote inflammation, pain and fever, and are necessary for blood clotting.
History of aspirinIn the 5 th century people used the bitterpowder extracted from a willow bark torelieve painsalicylic acid was used since 1874 as adrug• in 1897 Felix Hofmann formed acetylsalicylic acid by derivatizing one of the hydroxyl functional groups in salicylic acid with an acetyl group• Aspirin was patented in 1899 Felix Hofmann
Aspirin: Mechanism of Action Aspirin inhibits an enzyme, called COX which is responsible for the production of prostaglandines there are two forms : COX 1 and COX 2 COX 2COX 1 • induced ( normally not present in cells)• continuously stimulated by the body • built only in special cells (A549 lung• its concentration in the body remain cells)stable • used for signaling pain and• creates prostaglandins used for basic inflammationhouse keeping throughout body • produces prostaglandins for• prostaglandins stimulate normal body inflammatory responsefunctions such as stomach mucous • stimulated only as part of immuneproduction, regulation of gastric acid and responsekidney water excretion
Aspirin: Mechanism of ActionPlatelet Aggregation and Activation
Aspirin: Mechanism of Action Available Antiplatelet Agents
Indications• Aspirin used as,• Local analgetic effect• Antipyretic• Anti-inflammatory• Antiplatelet• Analgesic &Antipyretic• Rheumatic fever & heumatoid Arthritis• External applications Clinical aspirin resistance• Corns & calluses• Colon cancer • The inability of aspirin to protect a• Cardiovascular applications person from cardiovascular events such as heart attacks and strokes.• Prophylactically to decrease the incidence of ischemic attacks & unstable angina
AIM“Aspirin induced alterations in the Liver, Kidney and Reproductive System of female albino rat, Rattus rattus norvegicus”
Animals Selection• The present experiment were performed on mature female albino rat, Rattus rattus (n=20). The rats were obtained from the colony of the animal house of Barkatullah University, Bhopal. The rats were 15-16 weeks old, the body weight range 125 ± 5 g and acclimatized up to three weeks. The rats were maintained at suitable temperature of 23 ± 2ºC with an automatically controlled photo-period (12 hours light and 12 hours darkness) and the rats were fed with standard balanced pelleted diet with tap water "ad libitum". The changes in vaginal smear were also measured daily. The above experiment was performed during the month of November-10 to March- 11.
Chemical and Dose Selection• Aspirin tablets (Acetyl Salicylate, Ecosprin- 150) obtained from registered medical shop, Bhopal has been used for present experiment. The experimental animals were administered Aspirin orally at a dose level of 100 mg/Kg b.w./day through gavage.
Experimental designFor present investigation total 20 female rats were divided into 2 groups(ten each), these different groups and dosing patterns as follows. • Ist Group – The animals of this group were fed with normal diet, were called SET A or Control Group. • IInd Group – The animals of this group was fed with normal diet and received Aspirin by oral administration for 15 & 30 days, were called Treated Group. This group was divided into 2 sub groups as following. • Ist Sub Group – these groups of 5 animals was fed with normal diet and received Aspirin by oral administration for 15 days, was called SET B or Treated Group A. • IInd Sub Group – these groups of 5 animals was fed with normal diet and received Aspirin by oral administration for 30 days, was called SET C or Treated Group B.
Method Design• All animals were sacrificed after 24 hours of the last dose according to their dosing patterns i.e. 15 days treated animals killed 16th day and 30 days treated animals killed 31st day along with control animals. After sacrificing blood were collected through cardiac puncture and serum were prepared for the different parameters i.e. biochemical parameter etc. At the same time body organs viz. liver, kidney, ovary, oviduct, uterus were quickly removed, washed with physiological saline, dried (by filter paper), weighed (by physical precision balance). Then these organs either kept in Bovines fixative for histopathological study or tissue homogenate were prepared for enzymological study.
Parameter estimated Body Weight study: From 1-30 days of control and treated animals on different time interval i.e. initial, after 15 days and after 30 days. Hematological Study: Hb %, RBCs and WBCs count Enzymological Study:1) Acid Phosphatase (ACP): King and Kings Method (1954).2) Alkaline Phosphatase (ALP): King and Kings Method (1954).3) Glutamate Oxalate Transaminase (GOT): Reitman and Frankel Method (1957).4) Glutamate Pyruvate Transaminase (GPT): Reitman and Frankel Method (1957). Biochemical Study:1) Cholesterol Estimation: By Ferric Chloride and Sulphuric Acid Method (Henly, 1957).2) Protein Estimation: By Follin-Phenol Method (Lowry et al., 1951).3) Glucose Estimation: By Dinitro Salicylic Acid Method (Miller, 1959).4) Bilirubin Estimation: By Malloy and Evelyn Method (1937)5) Calcium Estimation: By Eriochrome Dye Method (Preston Smith et al., 1966). Histological study: Ovary, Oviduct, Uterus, Liver and Kidney. Estrus cycle study: From 1-30 days (daily) of control and treated animals.
Following phases are seen in rat vaginal smear•Estrus lasts up to 12 h and is indicated by the Cell types in vaginal smears of Rattus rattuspresence of large cornified cells in the vaginalsmear. Epithelial cells•Metestrus lasts 21 h and usually has many Х 80 Х 80neutrophils in the smear and scattered squamousepithelial cells.•Diestrus lasts up to 57 hours and there are Cornified cells Leucocytesabundant neutrophils and a few nucleated non-cornified epithelial cells.•Proestrus lasts 3-12 hours and has abundantnucleated non-cornified epithelial cells.
The regular study of vaginal smears of control and treated groups of rat (Rattus rattus). DAYS 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 C O N T D E M P D E M P D E M P D E M P D E M P D E M P D E M P D E R O LGROUP TS R E A E M P D E M P D E M P D E M P D E M P D D D D D D D D D D D T E DControl Group = This Group contains 5 animals which receive D, E, M and P = Showing estrus cycle phasesonly normal diet from 1 to 30 days. D = Diaestrus Phase E = Estrus PhaseTreated Group = This Group contains 5 animals which receive M = Metaestrus Phasenormal diet and aspirin (100mg/Kg b.w.) by oral administration P = Proestrus phasefrom 1 to 30 days.
DISCUSSION• Data of the present study indicate that, body weight of rats (n=5) were not significantly affected by oral administration of Aspirin (100mg/Kg b.w.) for 30 days. One could suggest that oral administration of Aspirin (0.05% w/v) do not affect body weight in rats (Ebuehi et al., 2007; Kwon et al., 1997).• Our present findings shows that, significant (<0.001) increase in the number of TC WBC in 30 days Aspirin treated rats. The auther suggested that, Aspirin increases the production of blood eosinophils (and possibly blood basophils) (Hirai et al., 2001).• After the administration of Aspirin (100mg/Kg b.w.) for 30 days do not significant altered TC RBC and Hb %. Accoding to auther, Aspirin had showed no effect or alteration on packed cell volume (Otimenyin, et al).• In our present study, the serum protein content were not significantly (p<0.01) reduced by oral administration of Aspirin. Ebuehi et al., (2007) suggested that, oral administration of Aspirin may inhibit protein synthesis or many prevent the uptake of amino acids into the tissues (such as in brain tissues).• The study of Aspirin on the serum Ca level suggest that, the Ca++ concentration significantly (p<0.01) decreased on the dose level 100mg/Kg b.w. of Aspirin. In addition to, Aspirin diet enhances calcium absorption for the improvement in hen’s egg shell percentage and thickness (Abdullah et al., 2010). Low levels of serum Calcium can be caused by low albumin or total protein levels (www.questdiagnostics.com, 2004).• The administration of Aspirin for 15 and 30 days on rats caused significant reduce the glucose and cholesterol level. Aspirin is known to inhibit glycogenesis and aminotransferase enzymes which may lead to decreased gluconeogenesis. The reduction of glucose and cholesterol levels which accompanied the reduction of corticosterone and T3 levels that consequently may affect its metabolism (Mohammed et al., 2010).
DISCUSSION• In our present study Aspirin significantly increases ACP and ALP content in Ovarian Utricular tissues and not significantly altered ACP and ALP content in Serum. The present investigation revealed that, Aspirin significantly increased serum GOT and GPT content in 15 days and 30 days treated rats at the dose level 100 mg/Kg (b.w.). As well as those patients, who are treated with NSAIDs, particularly with Aspirin, are at more danger of developing hepatotoxicity than others as well as the liver enzymes (ALT ,ALP, GOT and GPT) started to deteriorate gradually (Kawar et al, 2010).• In the present data indicates, Aspirin significantly reduces serum bilirubin content in rats compare to control rats. In the treatment of arthritis an anti-inflammatory drug were given (Aspirin level was 200 mg/kg/24hr) for two days found that, the bilirubin level were normal in patients (Kawar et al., 2010).• After the 15 and 30 days treatment of Aspirin (100mg/Kg b.w.) causes, significantly increase of serum Creatinine. Aspirin treated patients increase of serum Creatinine (Muhammad et al., 2010).• In the present investigation found that Aspirin induced toxicity on liver and Kidney both along with patho-histological changes at the higher dose level (100mg/Kg b.w./day). The results revealed that Aspirin made changes in antioxidant system even at therapeutic doses, made no changes in hepatic and renal pathology (Balog et al., 2000).
DISCUSSION• Our present findings show that, the morphological changes found in both ovaries and oviduct and uterus seen in the experimental groups may be due to the vascular properties of Aspirin. long-term administration of Aspirin for 30 days, developed an areas of hemorrhagic spots, at the site of uterine wall vascularization and this may be due to blood vessels weakness induced by the prolonged program of Aspirin- induced treatment (Schafer. et al., 1995).• Aspirin administration persistently arrest luteolysis (i.e. intact and functional corpora lutea), which has been confirmed by daily vaginal smear, that reflect a cellular picture of diestrous phase. Inhibitors of PGF2α and synthesis (indomethacin and acetylsalicylic acid) as the endogenous mediator of luteolysis, were shown to delay the regression of the corpora lutea and to prolong the luteal activity in pseudopregnant rats (Jezova et al., 1999).• structural ovarian changes seen: significant increase in the number of corpora lutea and a significant increase in the diameter of the granulose lutein cells, with absence of regressed corpora luteal when compared to the control animals.• The structural uterine changes by Aspirin namely: the significant decrease in the endometrial lining cell height, endometrial thickness and diameter of endometrial glands in both periods of Aspirin treated rat (15 and 30 days) (Chaffin et al., 2000).
SUMMARYThe data of the present study summarized as, the oral administration of Aspirin (100mg/Kg b.w.)do not significantly affected to the body weight, Hb%, TC RBC and protein content. As well assignificantly increases TC WBC. In addition, oral administration of Aspirin significantly inhibitedthe level of Glucose and cholesterol level. Therefore, it could be inferred the level of serum andtissues enzymes such as ACP, ALP, GOT and GPT. And also Aspirin significantly causedalteration in bilirubin and Creatinine concentration. Furthermore information about Aspirin is thatit also causes patho-histological changes in liver, kidney and reproductive organ (i.e. ovary,oviduct and uterus). It conclude from present study is that, the long term use of Aspirin at the dose of 100mg/kg b. w. orally is responsible for damage to female reproductive organs, if precautionary measures are not taken while using them. Structural damage to mammalian oviduct may be lead to infertility cause. So in selecting a safe drug, quantities and mode of usage must be strictly monitored to minimize the possibility of injurious complications. This can be achieved through public health education to make people aware of the hazardous effect of these compounds. It is therefore recommended that great precautions and supplements must be taken to minimize the harmful side effects of such chemicals especially to human and animals.
REFERENCES• Abdullah A. Mohammed (2010): Effect of acetyl salicylic acid (Aspirin) in drinking water on productive performance and blood characteristic of layer hens during heat stress. Int. J. Poult. Sci., vol. 9 (4), p382-385.• Acta Veterinaria, Turja A. Panteli, Drenka et al., (2010): Effects of Aspirin on the number of peripheral white blood cells and spleen eosinophils in guinea-pigs. Vol. 60, No. 2-3, p355-362.• Bonses, R.W. & Tausskay, H.H. (1945); J.Biol, Chem., 158, 581. Slot, C. (1965).• Scand J. Clin, Toro, G. & Ackermann P, (1975): Practical Clinical Chemistry, p154.• Buck ML, Grebe TA, Bond GR (1993): Toxic reaction to salicylate in a newborn infant: similarities to neonatal sepsis. J. Pediatr., 122:955-8.• Cheng Y, Austin SC, Rocca B, Koller BH, Coffman TM, Grosser T, Lawson JA, Fitz Gerald GA (2002): Role of prostacyclin in the cardiovascular response to thromboxane A2. Science, 19, 296(5567), p539-41.• Chignard M, Concalves de, Moraes VL, Lefort J, Meager A, Vargafting B (1996): Effect of cyclooxygenase inhibitors and modulators of cyclic AMP formation on lipopolysaccharide-inducedneutrophil infiltration in mouse lung. Br. J. Pharmacol., 117, 8.• Daud AN, De Silva KI, Deng J, Gamelli RL, Jones SB, Shanker R.(2003): Prostaglandin E2 mediates growth arrest in NFS-60 cells by down- regulating interleukin-6 receptor expression. Biochem. J, 370, Pt 1.• Decaterina R, Giannessi D, Bernini W (1985): Low-dose Aspirin in patient recovering from myocardial infarction: evidence for a selective inhibition of thromboxane-related platelet function. Eur. Heart. J., 6, p409-417.• Ebuehi, O.A.T., Q.C. Opara and A.I. Akinwande, (2007). Effects of oral administration of Aspirin and paracetamol on plasma and brain protection, tryptophane levels and monoamine oxidase activity in rats. Journal of Biological Sciences, 7 (8), p1468-1472.• Endo T, Yamamoto H, Tanaka S (1988): Effect of prostaglandins on progesterone production by human luteal cells & their ability of prostaglandin production. Nippon. Naibunpi Cakkai Zaschi, 20, 64(8), p687-97.• Ganong WF ((2003): Review of medical Physiology. A Lange medical book, 21 ed., McGraw-Hill.• Garcia-Rodriguez LA (2001): The effect of non steroidal anti-inflammatory drugs on the risk of coronary heart disease: fusion of clinical pharmacology and pharmacoepidemiologic data. Clin. Exp. Rheumatol, 19, (6 suppl. 25), p41-4.• Hall JE, Schoenfeld DH, Martin KA, et al (1992): Hypothalamic gonadotropin-releasing hormone secretion & folliclestimulating hormone dynamics during the luteal-follicular transition. J. Clin. Endocrinol. Metab., 74, p600-7.• Hall, D., et al. (1992): Counteraction of the Vasodilator Effects of Enalapril by Aspirin in Severe Heart Failure. J. Am. Coll. Cardiol., 20:1549- 1555).• J. M. Balog, 2 G. R. Huff, N. C. Rath, and W. E. Huff (2000): Effect of Dietary Aspirin on Ascites in Broilers Raised in a Hypobaric Chamber. Poultry Science 79:1101–1105.• Jezova M, Scsukova S, Vranova J, Kolena J (1999): Modulation of LH/hCG receptors & physical state of ovarian membranes in rat psuedopregnancy. Gen. Physiol. Biophys., 18(4), p347-56.• Lowry O.H., Rosebrough N.J., Farr A.L., Randal R.J. (1951): Protein measurement with the Folin phenol reagent. J. Bio. Chemistry, 193, 265.• Luigi Di Luigi, Laura Guidetti, Francesco Romanelli, Carlo Baldari, and Domenico Conte (2001): Aspirin, Exercise, And Pituitary Hormones. Official Journal of the American College of Sports Medicine, p2029-2035.• Marcia L. Buck, (2007): Use of Aspirin in Children with Cardiac Disease. Pediatric Pharmacotherapy,Vol. 13, Number 2.• McCannce KL, Huether SE (1994): Pathophysiology, the biologic bases for disease in adults and children. 2nd ed. Mosby year book, Inc., The menstrual cycle, p723.