Erythrocytes Membrane Alterations Reflecting Liver Damage in CCl₄-Induced Cirrhotic Rats: The Ameliorative Effect of Naltrexone
Cirrhosis is the consequence of chronic liver disease. Deleterious effects of oxidative stress on hepatocytes may be reflected in the erythrocyte membrane. Naltrexone (NTX) has been shown to attenuate hepatocellular injury in fibrotic animal models. The aim of this study was to investigate the progressive effect of CCl4 on the liver and whether the improvement of liver cirrhosis can be monitored through alterations in the erythrocyte membrane. In this study, 84 male Wistar rats were divided into 4 groups and received reagents (i.p.) as follows: 1- CCl₄, 2- NTX + CCl₄, 3- Mineral Oil (M), and 4- NTX + M. After 2, 6 and 8 weeks, the blood and liver tissue samples were collected. Plasma enzyme activities, the content of erythrocyte GSH and some membrane compositions, including protein carbonyl, protein sulfhydryl, and malondialdehyde were assessed. After 6 and 8 weeks, plasma enzyme activities and the content of protein carbonyl were higher in CCl4 group significantly, as compared to other groups (P<0.001). NTX significantly diminished protein carbonyl and plasma enzyme activities (P<0.001). GSH did not change until the 6th week. However, CCl4+NTX increased it significantly as compared to CCl₄ group (P<0.05). Protein sulfhydryl showed changes in NTX+CCl₄ group which indicated a significant increase in protein sulfhydryl content in a 6th week compared to CCl4 group (P<0.05). MDA did not show any significant alteration. CCl₄-induced cirrhosis is accompanied by increased content of oxidative stress markers, especially protein carbonyl of RBC membrane and plasma enzyme activities. This study shows that the progression of liver cirrhosis and the ameliorative effect of NTX can be followed through alterations of these markers.
Mokdad AA, Lopez AD, Shahraz S, Lozano R, Mokdad AH, Stanaway J, et al. Liver cirrhosis mortality in 187 countries between 1980 and 2010: a systematic analysis. BMC Medicine. 2014;12(1):145.
Williams R. Global challenges in liver disease. Hepatology. 2006;44(3):521-6.
Violi F, Leo R, Vezza E, Basili S, Cordova C, Balsano F, et al. Bleeding time in patients with cirrhosis: relation with degree of liver failure and clotting abnormalities. Journal of Hepatology. 1994;20(4):531-6.
Hernandez–Gea V, Toffanin S, Friedman SL, Llovet JM. Role of the microenvironment in the pathogenesis and treatment of hepatocellular carcinoma. Gastroenterology. 2013;144(3):512-27.
Shiraha H, Yamamoto K, Namba M. Human hepatocyte carcinogenesis (review). International Journal of Oncology. 2013;42(4):1133-8.
Coulouarn C, Clément B. Stellate cells and the development of liver cancer: Therapeutic potential of targeting the stroma. Journal of Hepatology. 2014.
Manns MP. Liver cirrhosis, transplantation and organ shortage. Deutsches Ärzteblatt International. 2013;110(6):83.
Patel K, Gordon SC, Jacobson I, Hézode C, Oh E, Smith KM, et al. Evaluation of a panel of non-invasive serum markers to differentiate mild from moderate-to-advanced liver fibrosis in chronic hepatitis C patients. Journal of Hepatology. 2004;41(6):935-42.
Marcellin P, Ziol M, Bedossa P, Douvin C, Poupon R, De Lédinghen V, et al. Non‐invasive assessment of liver fibrosis by stiffness measurement in patients with chronic hepatitis B. Liver International. 2009;29(2):242-7.
Halfon P, Munteanu M, Poynard T. FibroTest-ActiTest as a non-invasive marker of liver fibrosis. Gastroentérologie Clinique et Biologique. 2008;32(6):22-39.
Castéra L, Sebastiani G, Le Bail B, de Lédinghen V, Couzigou P, Alberti A. Prospective comparison of two algorithms combining non-invasive methods for staging liver fibrosis in chronic hepatitis C. Journal of Hepatology. 2010;52(2):191-8.
Geetha A, Lakshmi Priya M, Jeyachristy SA, Surendran R. Level of oxidative stress in the red blood cells of patients with liver cirrhosis. Indian Journal of Medical Research. 2007;126(3):204.
Rosa DPd, Bona S, Simonetto D, Zettler C, Marroni CA, Marroni NP. Melatonin protects the liver and erythrocytes against oxidative stress in cirrhotic rats. Arquivos de Gastroenterologia. 2010;47(1):72-8.
Grattagliano I, Giudetti A, Grattagliano V, Palmieri V, Gnoni G, Lapadula G, et al. Structural and oxidative modifications of erythrocyte ghosts in patients with primary biliary cirrhosis: relation with the disease stage and effect of bile acid treatment. European Journal of Clinical Investigation. 2003;33(10):868-74.
Moustafa AHA, Ali EM, Mohamed TM, Abdou HI. Oxidative stress and thyroid hormones in patients with liver diseases. European Journal of Internal Medicine. 2009;20(7):703-8.
Seki S, Kitada T, Yamada T, Sakaguchi H, Nakatani K, Wakasa K. In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fatty liver diseases. Journal of Hepatology. 2002;37(1):56-62.
Catalá A. Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chemistry and Physics of Lipids. 2009;157(1):1-11.
Barcia JM, Flores-Bellver M, Muriach M, Sancho-Pelluz J, Lopez-Malo D, Urdaneta AC, et al. Matching Diabetes and Alcoholism: Oxidative Stress, Inflammation, and Neurogenesis Are Commonly Involved. Mediators of Inflammation. 2014.
Moustafa AH, Ali EM, Mohamed TM, Abdou HI. Oxidative stress and thyroid hormones in patients with liver diseases. Eur J Intern Med. 2009 Nov;20(7):703-8.
Halliwell B, Gutteridge J. Oxygen free radicals and iron in relation to biology and medicine: some problems and concepts. Archives of Biochemistry and Biophysics. 1986;246(2):501-14.
Pandey KB, Rizvi SI. Markers of oxidative stress in erythrocytes and plasma during aging in humans. Oxidative Medicine and Cellular Longevity. 2010;3(1):2-12.
Yang X, Greenhaw J, Shi Q, Roberts DW, Hinson JA, Muskhelishvili L, et al. Mouse liver protein sulfhydryl depletion after acetaminophen exposure. Journal of Pharmacology and Experimental Therapeutics. 2013;344(1):286-94.
Ghezzo A, Visconti P, Abruzzo PM, Bolotta A, Ferreri C, Gobbi G, et al. Oxidative stress and erythrocyte membrane alterations in children with autism: correlation with clinical features. PLoS One. 2013;8(6):e66418.
Manjrekar A, Jisha V, Bag P, Adhikary B, Pai M, Hegde A, et al. Effect of Phyllanthus niruri Linn. treatment on liver, kidney and tests in CCl4 induced hepatotoxic rats. Indian Journal of Experimental Biology. 2008;46(7):514.
Ranawat L, Bhatt J, Patel J. Hepatoprotective activity of ethanolic extracts of bark of Zanthoxylum armatum DC in CCl 4 induced hepatic damage in rats. Journal of Ethnopharmacology. 2010;127(3):777-80.
Morakinyo A, OludareGO AA, Adegoke O. Protective effects of alpha lipoic acid on carbon tetrachloride-induced liver and kidney damage in rats. British Journal of Pharmacology and Toxicology. 2012;3:21-8.
Lee C, Park S, Kim YS, Kang SS, Kim JA, Lee SH, et al. Protective mechanism of glycyrrhizin on acute liver injury induced by carbon tetrachloride in mice. Biological and Pharmaceutical Bulletin. 2007;30(10):1898.
Wang T, Sun N-L, Zhang W-D, Li H-L, Lu G-C, Yuan B-J, et al. Protective effects of dehydrocavidine on carbon tetrachloride-induced acute hepatotoxicity in rats. Journal of Ethnopharmacology. 2008;117(2):300-8.
Fang H-L, Lai J-T, Lin W-C. Inhibitory effect of olive oil on fibrosis induced by carbon tetrachloride in rat liver. Clinical Nutrition. 2008;27(6):900-7.
Basu S. Carbon tetrachloride-induced lipid peroxidation: eicosanoid formation and their regulation by antioxidant nutrients. Toxicology. 2003;189(1):113-27.
Khan RA, Khan MR, Sahreen S. Evaluation of Launaea procumbens use in renal disorders: A rat model. Journal of Ethnopharmacology. 2010;128(2):452-61.
Javadi-Paydar M, Ghiassy B, Ebadian S, Rahimi N, Norouzi A, Dehpour AR. Nitric oxide mediates the beneficial effect of chronic naltrexone on cholestasis-induced memory impairment in male rats. Behavioural Pharmacology. 2013;24(3):195-206.
Kiani S, Ebrahimkhani MR, Shariftabrizi A, Doratotaj B, Payabvash S, Riazi K, et al. Opioid system blockade decreases collagenase activity and improves liver injury in a rat model of cholestasis. Journal of Gastroenterology and Hepatology. 2007;22(3):406-13.
De Minicis S, Candelaresi C, Marzioni M, Saccomano S, Roskams T, Casini A, et al. Role of endogenous opioids in modulating HSC activity in vitro and liver fibrosis in vivo. Gut. 2008;57(3):352-64.
Marzioni M, Alpini G, Saccomanno S, De Minicis S, Glaser S, Francis H, et al. Endogenous opioids modulate the growth of the biliary tree in the course of cholestasis. Gastroenterology. 2006;130(6):1831-47.
Almansa I, Barcia JM, López-Pedrajas R, Muriach M, Miranda M, Romero FJ. Naltrexone Reverses Ethanol-Induced Rat Hippocampal and Serum Oxidative Damage. Oxidative Medicine and Cellular Longevity. 2013;2013.
Restellini S, Alaei M, Matthes T, Kherad O, Moschetta A, Spahr L. Effect of hydrosoluble vitamin E on erythrocyte membrane lipid composition in patients with advanced cirrhosis: An open‐label pilot trial. Hepatology Research. 2015.
Dodge JT, Mitchell C, Hanahan DJ. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Archives of Biochemistry and Biophysics. 1963;100(1):119-30.
Gornall AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J biol Chem. 1949;177(2):751-66.
Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz A-G, et al. Determination of carbonyl content in oxidatively modified proteins. Methods in Enzymology. 1990;186:464.
Habeeb A. Reaction of protein sulfhydryl groups with Ellman's reagent. Methods Enzymol. 1972;25:457-64.
Drabkin D, Austin J. Spectrophotometric studies: Spectrophotometric constants for common hemoglobin derivatives in human, dog and rabbit blood. J Biol Chem. 1932;98(719):33.
Buege JA, Aust SD. Microsomal lipid peroxidation. Methods in Enzymology. 1978;52:302.
Halliwell B, Gutteridge J. Oxygen radicals and the nervous system. Trends in Neurosciences. 1985;8:22-6.
de Andrade Belo MA, Soares VE, de Souza LM, da Rosa Sobreira MF, Cassol DMS, Toma SB. Hepatoprotective treatment attenuates oxidative damages induced by carbon tetrachloride in rats. Experimental and Toxicologic Pathology. 2012;64(3):155-65.
Selvi S, Umadevi P, Suja S, Sridhar K, Chinnaswamy P. Inhibition of in vitro lipid peroxidation (LPO) evoked by Calocybe indica (Milky Mushroom). Ancient Science of Life. 2006;26(1-2):42.
Choi J, Ou J-HJ. Mechanisms of liver injury. III. Oxidative stress in the pathogenesis of hepatitis C virus. American Journal of Physiology-Gastrointestinal and Liver Physiology. 2006;290(5):G847-G51.
Doustimotlagh AH, Dehpour AR, Nourbakhsh M, Golestani A. Alteration in Membrane Protein, Antioxidant Status and Hexokinase Activity in Erythrocytes of CCl4-Induced Cirrhotic Rats. Acta Medica Iranica. 2014.
Faramarzi N, Abbasi A, Tavangar SM, Mazouchi M, Dehpour AR. Opioid receptor antagonist promotes angiogenesis in bile duct ligated rats. Journal of Gastroenterology and Hepatology. 2009;24(7):1226-9.
Brewer KL, Troendle MM, Pekman L, Meggs WJ. Naltrexone prevents delayed encephalopathy in rats poisoned with the sarin analogue diisopropylflurophosphate. The American Journal of Emergency Medicine. 2013;31(4):676-9.
Thomas CP, Wallack SS, Lee S, McCarty D, Swift R. Research to practice: adoption of naltrexone in alcoholism treatment. Journal of Substance Abuse Treatment. 2003;24(1):1-11.
Day SA, Lakner AM, Moore CC, Yen M-H, Clemens MG, Wu ES, et al. Opioid-like compound exerts anti-fibrotic activity via decreased hepatic stellate cell activation and inflammation. Biochemical Pharmacology. 2011;81(8):996-1003.
Zhang YT, Zheng QS, Pan J, Zheng RL. Oxidative damage of biomolecules in mouse liver induced by morphine and protected by antioxidants. Basic & Clinical Pharmacology & Toxicology. 2004;95(2):53-8.
Ebrahimkhani MR, Kiani S, Oakley F, Kendall T, Shariftabrizi A, Tavangar SM, et al. Naltrexone, an opioid receptor antagonist, attenuates liver fibrosis in bile duct ligated rats. Gut. 2006;55(11):1606-16.
Tirmenstein MA, Nelson SD. Acetaminophen-induced oxidation of protein thiols. Contribution of impaired thiol-metabolizing enzymes and the breakdown of adenine nucleotides. Journal of Biological Chemistry. 1990;265(6):3059-65.
Andringa KK, Bajt ML, Jaeschke H, Bailey SM. Mitochondrial protein thiol modifications in acetaminophen hepatotoxicity: effect on HMG-CoA synthase. Toxicology Letters. 2008;177(3):188-97.
Smith CV, Mitchell JR. Acetaminophen hepatotoxicity invivo is not accompanied by oxidant stress. Biochemical and Biophysical Research Communications. 1985;133(1):329-36.
|Issue||Vol 54, No 10 (2016)|
|Cirrhosis Carbon tetrachloride Naltrexone Oxidative stress|
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