|Year : 2015 | Volume
| Issue : 1 | Page : 21-27
Amelioration of cisplatin-induced hepatotoxicity by statins in rats
RA Maheshwari1, GU Sailor1, AK Sen1, R Balaraman2
1 Assistant Professor, Department of Pharmacy, Sumandeep Vidyapeeth, Piparia, Vadodara-391 760, Gujarat, India
2 Professor, Department of Pharmacy, Sumandeep Vidyapeeth, Piparia, Vadodara-391 760, Gujarat, India
|Date of Web Publication||3-Aug-2018|
R A Maheshwari
Assistant Professor, Department of Pharmacy, Sumandeep Vidyapeeth, Piparia, Vadodara-391 760, Gujarat
Source of Support: None, Conflict of Interest: None
Aim: This study was aimed to investigate the effect of simvastatin (SIM) and rosuvastatin (ROS) on the extent of tissue damage in cisplatin (CIS) induced hepatotoxicity.
Methodology: Hepatotoxicity was induced in rats with single intraperitoneal injection of 7 mg/kg cisplatin. Group 1 received 0.5% sodium carboxy methyl cellulose, group 2 and 3 received SIM and ROS, respectively, and group 4 was injected single dose of CIS (7 mg/kg, i.p.).Group 5 and 6 were treated with SIM (10 mg/kg, p.o.) and ROS (10 mg/kg, p.o.) daily from 5 days before to 5 day after intraperitoneal administration of CIS, respectively. Liver function tests like AST, ALT and Total bilirubin, and markers of oxidative stress such as liver malondialdehyde (MDA) level, superoxide dismutase (SOD), catalase (CAT) activities and reduced glutathione (GSH) were measured. All tissues were investigated for histopathological changes.
Results: CIS treated rats showed a significant increase in AST, ALT and total bilirubin. Moreover, cisplatin caused liver damage with a higher MDA level, depletion of SOD, CAT activity and GSH. SIM and ROS ameliorate CIS induced liver damage due to improvement in liver function, oxidative stress, and histological alteration.
Conclusions: These finding suggests that simvastatin and rosuvastatin may have a protective effect against cisplatin-induced liver damage via amelioration of lipid peroxidation as well as due to improvement of liver function.
Keywords: Cisplatin, Simvastatin, Rosuvastatin, Hepatotoxicity
|How to cite this article:|
Maheshwari R A, Sailor G U, Sen A K, Balaraman R. Amelioration of cisplatin-induced hepatotoxicity by statins in rats. J Integr Health Sci 2015;3:21-7
|How to cite this URL:|
Maheshwari R A, Sailor G U, Sen A K, Balaraman R. Amelioration of cisplatin-induced hepatotoxicity by statins in rats. J Integr Health Sci [serial online] 2015 [cited 2020 May 28];3:21-7. Available from: http://www.jihs.in/text.asp?2015/3/1/21/238515
| Introduction|| |
Statins are widely used clinically for lowering hypercholesterolemia because of their inhibitory effect on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the enzyme that catalyzes the rate-limiting step of the cholesterol synthesis in the liver and other tissues. Statins have beneficial changes in other lipid fractions, along with significant effects on numerous measures of inflammation, immunity, oxidative stress, thrombosis, vascular and renal function.,,,,,, Pleiotropic effects of statins have also been implicated in reductions in heart failure, myocardial ischemia, myocardial infarction, inflammatory responses to sepsis and with neuroprotection. Small randomized controlled clinical trials have also demonstrated beneficial effects in Non-cardiovascular inflammatory disorders, such as rheumatoid arthritis and multiple sclerosis.
Cis-dichlorodiammine platinum (II), or cisplatin (CIS), is currently among the most widely used agent in the chemotherapy of cancer.,, The chief limit to its greater efficacy is its nephrotoxicity. Although cisplatin-induced nephrotoxicity has been very well documented in clinical oncology, hepatotoxicity has been rarely characterized, and is less studied. It is well known fact that cisplatin is significantly taken up by human liver and that high dose of the drug produces hepatotoxicity. Generally liver toxicity of cisplatin is characterized by mild to moderate elevation of serum transaminases and less frequently, by a mild elevation of serum alkaline phophatase, lactate dehydrogenase (LDH), bilirubin and c- glutamyltranspeptidase levels. However, the mechanism of cisplatin induced liver damage is not well understood, although apoptotic lesions seem to characterize the damaged liver parenchyma. In a study by Koc et al. the histological analysis of liver tissue revealed cytoplasmic changes especially around cells of central vein and hepatocellular vacuolization and sinusoidal dilatations.
Therefore it was thought worthwhile to study the protective effect of simvastatin or rosuvastatin in cisplatin-induced hepatotoxicity and evaluate the possibility of using statins in preventing certain toxicity related to anticancer drug and some disease like ulcerative colitis and kidney damage where current therapy has limitation.
| Methodology|| |
Drugs and Chemicals
Simvastatin (SIM) and Rosuvastatin (ROS) were obtained as a gift sample from ZydusCadila, Ahmedabad, India. Cisplatin (CIS) was obtained from VHB limited, Mumbai, India. All other chemicals and reagents used in the study were of analytical grade.
All experimental protocols described in present study were approved by the Institutional Animal Ethics Committee (IAEC) and Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA). The experiment was carried out on healthy adult female Wistar rats weighing 200-250g. Rats were housed in polypropylene cages, maintained under standardized condition (12-h light/dark cycle, 24°C, 35 to 60% humidity) and provided ad libitium to palleted CHAKKAN diet (Nav Maharashtra Oil Mills Pvt. Ltd., Pune) and purified drinking water.
Cisplatin-induced liver damage
The rats were divided into six groups of six animals each.
Group 1: Normal control rats (1 ml/ kg of 0.5% sodium CMC, p.o.)
Group 2: SIM control rats (10 mg/kg/ml, p.o.)
Group 3: ROS control rats (10 mg/kg/ml, p.o.)
Group 4: CIS control rats (7 mg/kg/ml, i.p.),
Group 5: SIM (10 mg/kg/ml, p.o) was given daily from 5 days prior to 5 day after intraperitoneal administration of CIS.
Group 6: ROS (10 mg/kg, p.o) was given daily from 5 days before to 5 day after intraperitoneal administration of CIS
All the aforementioned treatments were given orally for 10 days and cisplatin was given intraperitoneally. At the end of treatment (10th day), blood samples were withdrawn from retro-orbital plexus under light ether anesthesia without any anticoagulant and allowed for 10 minutes to clot at room temperature. It was centrifuged at 2500 rpm for 20 minutes for serum separation. The serum obtained was kept at 4°C until further biochemical estimation. Total bilirubin (TB), serum alanine aminotransferase (ALT) and serum aspartate aminotransferase (AST) were estimated from serum sample using standard assay kits of SPAN Diagnostic Ltd., India.
Estimation of biomarkers of Oxidative stress
At end of the treatments, all the animals were sacrificed by cervical dislocation. Liver was dissected out and kept in cold conditions. The tissues were cross chopped with surgical scalpel into fine slices in chilled 0.25 M sucrose, quickly blotted on a filter paper. The tissue were minced and homogenized in 10 mMTris-HCl buffer, pH 7.4 (10%w/v) with 25 strokes of tight Teflon pestle of glass homogenizer at a speed of 2500 rpm. The clear supernatant was used for the assay of malondialdehyde  (MDA),activity of endogenous antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH).
Histopathology of liver
The liver tissue was rapidly dissected out, washed with saline immediately and fixed in 10% buffered formaldehyde. Paraffin-embedded specimens were cut into 5 μm-thick sections and stained with hematoxylin and eosin (H&E). The sections were examined under the light microscope (Olympus BX10, Japan) for histopathological changes and photomicrographs (Olympus DP12 camera, Japan) were taken. The pathologist performing histopathological evaluation was blinded to the treatment as assignment of different study groups.
All of the data are expressed as mean ± SEM. Statistical significance between more than two groups was tested using one-way ANOVA followed by the Bonferroni multiple comparisons test as appropriate using a computer-based fitting program (Prism, Graph pad 5). Differences were considered to be statistically significant when P < 0.05.
| Results|| |
Effect of statins on body weight in cisplatin- induced hepatotoxicity
The effects of SIM and ROS on body weight of rat were shown in [Table 1]. Body weight of CIS control rats were significantly (P < 0.001) decreased on day 10 compared to normal control rats, while treatment with SIM and ROS did not show any significant changes in body weight during cisplatin treatment.
|Table 1: Effect of SIM and ROS on body weight in cisplatin-induced hepatotoxicity|
Click here to view
Effect of statins AST, ALT and total bilirubin in cisplatin-induced hepatotoxicity
Administration of single injection of cisplatin (7 mg/kg, i.p.) caused a significant (P < 0.001) increase in AST, ALT and total bilirubin levels as compared to normal control animals. The treatment with SIM and ROS showed a significantdecrease in the levels of AST (P < 0.01; P < 0.001), ALT (P < 0.05; P < 0.01) and TB (P < 0.001), respectively as compared to CIS control rats [Table 2].
|Table 2: Effect of SIM and ROS on AST, ALT and total bilirubin in cisplatin-induced hepatotoxicity in rats|
Click here to view
Effect of statins on markers of oxidative stress
The content of MDA, end product of lipid peroxidation and marker of oxidative stress was significantly (P < 0.001) increased in liver tissue of CIS control rats as compared to normal rats. There was a significant (P < 0.001) decrease in the levels of GSH, an endogenous antioxidant and antiperoxidative enzymes (SOD and CAT) in liver tissue as compared to normal control group. The treatment with SIM showed a significant increase in SOD (P < 0.05), CAT (P < 0.01), GSH (P < 0.01) levels and a significant decrease in MDA (P < 0.05) levels as compared to CIS-treated group. The treatment with ROS showed a significant increase in SOD (P < 0.01), CAT (P < 0.001), GSH (P < 0.001) levels and significant decrease in MDA (P < 0.01) level as compared to CIS control rats [Table 3].
|Table 3: Effect of SIM and ROS on hepatic levels of SOD, CAT, GSH and MDA in cisplatin induced hepatotoxicity in rats|
Click here to view
The hepatoprotective effect of SIM and ROS were confirmed by histopathological examination of liver tissue. The liver sections of normal control, SIM control and ROS control rats showed no significant change, suggesting no direct effect of SIM and ROS on Liver. The normal liver architecture was disturbed in rats treated with cisplatin (CIS), showed severe activation of Kupffer cells, degenerated hepatocytes and moderate enlargement of sinusoids compared normal control group. SIM and ROS treatment partially preserved the hepatocytes degeneration, sinusoids and Kupffer cells was near to control morphology [Figure 1].
|Figure 1: Photomicrograph of liver of following groups.|
Control group: normal histological appearance of liver (A); SIM control group and ROS control group shows normal histological appearance (B) and (C); Cisplatin group: degenerated hepatocytes (arrow head), dilatation and vascular congestion in sinusoids (v) increased number of activated Kupffer cells (arrow), and inflammatory cell infiltration in portal triad region (*) in liver (D); SIM+CIS group: mild dilatation and vascular congestion in sinusoids (v), degenerated hepatocytes (arrow head), and activated Kupffer cells (arrow) in liver (E); ROS+CIS group: mild dilatation and vascular congestion in sinusoids (v), degenerated hepatocytes (arrow head), and activated Kupffer cells (arrow) in liver (F)
Click here to view
| Discussion|| |
The present study was aimed to investigate possibility of SIM and ROS to protect hepatotoxicity associated with anticancer drugs. The results of the present study reveal that treatment with SIM and ROS markedly ameliorate cisplatin-induced liver damage, as shown in microscopic examination and biochemical parameters. Cisplatin, one of the most active cytotoxic agents against cancer, has several toxicities. Liver damage is one of the toxic effect which occured during the high dose of the treatment,.,
In the present study, cisplatin administration caused severe liver damage characterized by severe activation of Kupffer cells, degenerated hepatocytes and moderate enlargement of sinusoids. Histopathological examination was also supplemented by increase in enzymes such as AST, ALT and TB. The ability of cisplatin to cause alterations in the activity of these enzymes could be a secondary event following cisplatin-induced liver damage with the consequent leakage from hepatocytes. The treatment with SIM and ROS reduced elevated levels of serum AST, ALT and TB indicating the protective effect of the liver.
The results of present study also revealed that a single dose of cisplatin cause decreased activity of SOD, CAT and GSH level, while increased in MDA level. This effect may be a secondary event following the cisplatin-induced increase in free radical generation and/or decrease in lipid peroxidation protecting enzymes. Cisplatin can cause the generation of oxygen free radicals, such as hydrogen peroxide, superoxide anions and hydroxyl radicals. The hydroxyl radical is capable of abstracting a hydrogen atom from polyunsaturated fatty acids in membrane lipids to initiate lipid peroxidation. These radicals can evoke extensive tissue damage, reacting with macromolecules, such as membrane lipids, proteins and nucleic acids. Moreover, depletion of glutathione may contribute to cisplatin-induced lipid peroxidation.
Catalase (CAT) is an enzymatic antioxidant widely distributed in all animal tissues. Superoxide dismutase (SOD) acts as a cellular defense element against potentially harmful effects of superoxide ions by catalyzing the dismutation of these ions. These results indicate that SOD plays a defensive role in cisplatin-induced hepatotoxic rats. Catalase decomposes hydrogen peroxide and protects the tissue from highly reactive hydroxyl radicals. Therefore reduction in the activity of these enzymes may result in a number of deleterious effects due to the accumulation of superoxide radicals and hydrogen peroxide. Administration of SIM and ROS restored the activities of SOD and CAT in cisplatin intoxicated rats. Our results revealed that SIM and ROS prevented excessive free radicals accumulation and protected the liver from cisplatin induced hepatotoxicity. Cisplatin intoxication produced signification depletion of GSH and imbalance of GSH/GSSG (glutathione disulfide) ratio. The reduced form of GSH becomes readily oxidized to GSSG on interacting with free radicals. Excessive production of free radicals resulted in the oxidative stress, which leads to damage to biomolecules e.g. lipids, and can induce lipid peroxidation in-vivo. In present study, the decrease level of GSH was associated with enhanced lipid peroxidation in cisplatin intoxicated rats. Administration of SIM and ROS restored the depleted glutathione levels, thus SIM and ROS may act by inducing the detoxifying enzymes and these enzymes might detoxify the free radicals produced following cisplatin intoxication.
Malondialdehyde (MDA) is an end-product of the breakdown of polyunsaturated fatty acids and related esters, and its formation is an index of lipid peroxidation in many organ homogenate. Treatment with SIM and ROS also decrease in MDA levels which is responsible for the formation of lipid peroxidation. Therefore, SIM and ROS reduced the levels of MDA indicating protective antioxidant effect on the animals treated with cisplatin.
| Conclusion|| |
The present study demonstrated that SIM and ROS have a significant hepatoprotective effect against CIS induced hepatotoxicity in rats. The enhanced levels of antioxidant enzymes and reduced amount of lipid peroxidation are suggested to be the main mechanisms of SIM and ROSin preventing the development of liver damage induced by CIS. This data along with histopathological examination also justified the result. Finally it was concluded that use of statins can prevent liver toxicity associated with anticancer drugs like cisplatin.
| References|| |
Corsini A, Maggi FM, Catapano AL. Pharmacology of competitive inhibitors of HMG- CoA reductase. Pharmacological research. 1995 Dec 31;31(1):9-27.
Davignon J. Beneficial cardiovascular pleiotropic effects of statins. Circulation. 2004; 109 (23 suppl 1): III 39-43.
Nissen SE. High-dose statins in acute coronary syndromes: not just lipid levels. Jama. 2004 Sep 15;292(11):1365-7.
Ray KK, Cannon CP. The potential relevance of the multiple lipid-independent (pleiotropic) effects of statins in the management of acute coronary syndromes. Journal of the American College of Cardiology. 2005 Oct 18;46(8):1425-33.
Douglas K, O’Malley PG, Jackson JL. Meta-analysis: the effect of statins on albuminuria. Annals of internal medicine. 2006 Jul 18;145(2):117-24.
Balk EM, Karas RH, Jordan HS, Kupelnick B, Chew P, Lau J. Effects of statins on vascular structure and function: a systematic review. The American journal of medicine. 2004 Nov 15;117(10):775-90.
Rosenson RS, Tangney CC. Antiatherothrombotic properties of statins: implications for cardiovascular event reduction. Jama. 1998 May 27;279(20):1643-50.
Mason RP, Walter MF, Jacob RF. Effects of HMG-CoA reductase inhibitors on endothelial function role of microdomains and oxidative stress. Circulation. 2004 Jun 1;109(21 suppl 1):II-34.
Khush KK, Waters DD, Bittner V, Deedwania PC, Kastelein JJ, Lewis SJ, Wenger NK. Effect of high-dose atorvastatin on hospitalizations for heart failure subgroup analysis of the Treating to New Targets (TNT) study. Circulation. 2007 Feb 6;115(5):576-83.
Deedwania P, Stone PH, Merz CN, Cosin- Aguilar J, Koylan N, Luo D, Ouyang P, Piotrowicz R, Schenck-Gustafsson K, Sellier P, Stein JH. Effects of Intensive Versus Moderate Lipid-Lowering Therapy on Myocardial Ischemia in Older Patients With Coronary Heart Disease Results of the Study Assessing Goals in the Elderly (SAGE). Circulation. 2007 Feb 13;115(6):700-7.
Pasceri V, Patti G, Nusca A, Pristipino C, Richichi G, Di Sciascio G, ARMYDA Investigators. Randomized trial of atorvastatin for reduction of myocardial damage during coronary intervention results from the ARMYDA (Atorvastatin for Reduction of Myocardial Damage during Angioplasty) Study. Circulation. 2004 Aug 10;110(6):674-8.
Terblanche M, Almog Y, Rosenson RS, Smith TS, Hackam DG. Statins and sepsis: multiple modifications at multiple levels. The Lancet infectious diseases. 2007 May 31;7(5):358-68.
Rajanikant GK, Zemke D, Kassab M, Majid A. The therapeutic potential of statins in neurological disorders. Current medicinal chemistry. 2007 Jan 1;14(1):103-12.
McCarey DW, McInnes IB, Madhok R, Hampson R, Scherbakova O, Ford I, Capell HA, Sattar N. Trial of Atorvastatin in Rheumatoid Arthritis (TARA): double-blind, randomised placebo- controlled trial. The Lancet. 2004 Jun 19;363(9426):2015-21.
Vollmer T, Key L, Durkalski V, Tyor W, Corboy J, Markovic-Plese S, Preiningerova J, Rizzo M, Singh I. Oral SIM treatment in relapsing- remitting multiple sclerosis. Lancet. 2004; 363: 1607–1608.
Vollmer T, Key L, Durkalski V, Tyor W, Corboy J, Markovic-Plese S, Preiningerova J, Rizzo M, Singh I. Oral simvastatin treatment in relapsing- remitting multiple sclerosis. The Lancet. 2004 May 15;363(9421):1607-8.
Einhorn LH. Curing metastatic testicular cancer. Proceedings of the National Academy of Sciences. 2002 Apr 2;99(7):4592-5.
Borch RF. The platinum antitumor drugs. In: Powis G, Prough RA (Eds.), Metabolism and Action of Drugs. London: Taylor & Francis; 1987, p.193.
Rozencweig M, Von Hoff DD, Slavik M, Muggia FM. Cis-diamminedichloroplatinum (II): a new anticancer drug. Annals of Internal Medicine. 1977 Jun 1;86(6):803-12.
Hesketh MA, Twaddell T, Finn A. A possible role for cisplatin (DDP) in the transient hepatic enzyme elevation noted after ondansetron administration. InProc Am Assoc Clin Oncol 1990 (Vol. 9, No. 323, p. b28).
Cavalli F, Tschopp L, Sonntag RW, Zimmermann A. Cisplatin-induced hepatic toxicity. Cancer Treat Rep. 1978;62:2125-6.
Koc A, Duru M, Ciralik H, Akcan R, Sogut S. Protective agent, erdosteine, against cisplatin- induced hepatic oxidant injury in rats. Molecular and cellular biochemistry. 2005 Oct 1;278(1- 2):79-84.
Upaganlawar AB, Rathod SP, Balaraman R. Effect of coenzyme Q10 on oxidative stress and membrane bound enzyme in cisplatin induced nephrotoxicity. Advance Pharmacology and Toxicology.2008; 9(3): 95-100.
İşeri S, Ercan F, Gedik N, Yüksel M, Alican I. Simvastatin attenuates cisplatin-induced kidney and liver damage in rats. Toxicology. 2007 Feb 12;230(2):256-64.
Slater TF, Sawyer BC. The stimulatory effects of carbon tetrachloride and other halogenoalkanes on peroxidative reactions in rat liver fractions in vitro. General features of the systems used. Biochemical Journal. 1971 Aug 1;123(5):805-14.
Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. Journal of Biological chemistry. 1972 May 25;247(10):3170-5.
Hugo EB. Oxidoreductases acting on groups other than CHOH: catalase. In: Colowick SP, Kaplan NO, Packer L, editors. Methods in Enzymology, vol. 105. London: Academic Press 1984; 121– 125.
Moron MS, Depierre JW, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochimica et Biophysica Acta (BBA)- General Subjects. 1979 Jan 4;582(1):67-78.
Zicca A, Cafaggi S, Mariggiò MA, Vannozzi MO, Ottone M, Bocchini V, Caviglioli G, Viale M. Reduction of cisplatin hepatotoxicity by procainamide hydrochloride in rats. European journal of pharmacology. 2002 May 10;442(3):265-72.
Kim SH, Hong KO, Chung WY, Hwang JK, Park KK. Abrogation of cisplatin-induced hepatotoxicity in mice by xanthorrhizol is related to its effect on the regulation of gene transcription. Toxicology and applied pharmacology. 2004 May 1;196(3):346-55.
Antunes LM, Darin JD, Maria de Lourdes PB. Effects of the antioxidants curcumin or selenium on cisplatin-induced nephrotoxicity and lipid peroxidation in rats. Pharmacological Research. 2001 Feb 28;43(2):145-50.
Halliwell B, Gutteridge JMC. Free radicals in biology and medicine, 2nd Ed., Oxford: Clarendon press;1989, p.86.
Chance B, Greenstein DS, Roughton FJ. The mechanism of catalase action. I. Steady-state analysis. Archives of Biochemistry and Biophysics. 1952 Jun 1;37(2):301-21.
Gupta M, Kumar RS. Hepatoprotective effects and antioxidant role of Caesalpinia bonducella on paracetamol-induced hepatic damage in rats. Natural product sciences. 2003;9(3):186-91.
Ip SP, Poon MK, Che CT, Ng KH, Kong YC, Ko KM. Schisandrin B protects against carbon tetrachloride toxicity by enhancing the mitochondrial glutathione redox status in mouse liver. Free Radical Biology and Medicine. 1996 Dec 31;21(5):709-12.
Sinclair AJ, Barnett AH, Lunie J. Free radical and autooxidant systems in health and disease. J Appl Med. 1991; 17: 409.
Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutrition, metabolism and cardiovascular diseases. 2005 Aug 31;15(4):316-28.
[Table 1], [Table 2], [Table 3]