Acta Medica Iranica 2014. 52(11):795-803.

Alteration in membrane protein, antioxidant status and hexokinase activity in erythrocytes of CCl4- induced cirrhotic rats.
Amir Hossein Doustimotlagh, Ahmad Reza Dehpour, Mitra Nourbakhsh, Abolfazl Golestani

Abstract


Several studies have shown that hepatocyte membrane composition changes in patients with cholestasis and cirrhosis. These alterations that are because of intracellular oxidative stress are supposed to be reflected in erythrocyte membrane. The aim of this study was to investigate the modification of erythrocyte membrane along with hexokinase and antioxidant enzymes during development of cirrhosis. Cirrhosis was induced by intraperitoneal injection of CCl4 in male Wistar rats. The test groups were: baseline, cholestatic, early cirrhotic and advanced cirrhotic along with an equal number of sham-control animals. The erythrocyte membrane modifications (protein sulfhydryl, protein carbonyl, and lipid peroxidation), as well as NO metabolites, were assessed. Activities of GPX, CAT, SOD and HK were also measured. Protein sulfhydryl content of the erythrocyte membrane (after 2, 6 and 10 weeks of injection) had significant progressive decrease. In contrast, protein carbonyls were remarkably increased 2 weeks after injection but significantly decreased after 6 weeks and returned to normal levels after 10 weeks. No significant difference in erythrocyte HK activity or MDA content was observed. Test groups showed significantly lower erythrocyte GPx activity after six weeks and CAT and SOD activities along with NO metabolites content after two weeks (P<0.05). This study indicates that the progression of cirrhosis is accompanied by alterations in antioxidant enzyme and decreased NO metabolites. Protein carbonyl alteration occurs in the early stages of cirrhosis while protein sulfhydryl alterations have a progressive decrease in advanced cirrhosis.

Keywords


Rats; Erythrocytes; Cholestasis; Liver cirrhosis; Oxidative stress; Protein carbonylation

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References


Hwang TL, Chen CY. Gender Different Response to Immunonutrition in Liver Cirrhosis with Sepsis in Rats. Nutrients 2012;4(3):231-42.

Wallace K, Burt A, Wright M. Liver fibrosis. Biochem J 2008;411(1):1-18.

Masaad AA, Aziza AE, Nabila SH, et al. Hepatoprotective Effect of Sarcophine Isolated from Soft Coral (Sarcophyton glaucum) in Rats. Global Veterinaria 2012;8(3):244-53.

El-Serag HB, Rudolph KL. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 2007;132(7):2557-76.

Ha HL, Shin HJ, Feitelson MA, et al. Oxidative stress and antioxidants in hepatic pathogenesis. World J Gastroenterol 2010;16(48):6035-43.

Gate L, Paul J, Ba GN, et al. Oxidative stress induced in pathologies: the role of antioxidants. Biomed Pharmacother 1999;53(4):169-80.

Moustafa AH, Ali EM, Mohamed TM, et al. Oxidative stress and thyroid hormones in patients withliver diseases. Eur J Intern Med 2009;20(7):703-8.

Seki S, Kitada T, Yamada T, et al. In situ detection of lipid peroxidation and oxidative DNA damage in non-alcoholic fattyliver diseases. J Hepatol 2002;37(1):56-62.

Pandey KB, Rizvi SI. Markers of oxidative stress in erythrocytes and plasma during aging in humans. Oxid Med Cell Longev 2010;3(1):2-12.

Rodriguez-Vilarrupla A, Bosch J, Garcia-Pagan JC. Potential role of antioxidants in the treatment of portal hypertension. J Hepatol 2007;46(2):193-7.

Bosch J, Garcia-Pagan JC. Complications of cirrhosis. I. Portal hypertension. J Hepatol 2000;32(1 Suppl):141-56.

Wiest R, Groszmann RJ. The paradox of nitric oxide in cirrhosis and portal hypertension: too much, not enough. Hepatology 2002;35(2):478-91.

Domenicali M, Caraceni P, Giannone F, et al. A novel model of CCl4-induced cirrhosis with ascites in the mouse.J Hepatol 2009;51(6):991-9.

Galicia-Moreno M, Rodríguez-Rivera A, Reyes-Gordillo K, et al. N-acetylcysteine prevents carbon tetrachlorideinduced liver cirrhosis: role of liver transforming growth factor-beta and oxidative stress. Eur J Gastroenterol Hepatol 2009;21(8):908-14.

Ghasemi A, Hedayati M, Biabani H. Protein precipitation methods evaluated for determination of serum nitric oxide end products by the Griess assay. J Med Sci Res 2007;2(1):43-6.

Benzie IFF, Strain JJ. The ferric reducing ability of plasma(FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 1996;239(1):70-6.

Parpart AK, Lorenz PB, Parpart ER, et al. The osmotic resistance (fragility) of human red cells. J Clin Invest 1947;26(4):636-40.

Dodge JT, Mitchell C, Hanahan DJ. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys 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.

Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1978;52:302-10.

Levine RL, Garland D, Oliver CN, et al. Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 1990;186:464-78

Habeeb A. Reaction of protein sulfhydryl groups with Ellman's reagent. Methods Enzymol 1972;25:457-64.

Drabkin DL, Austin JH. Spectrophotometric studies: Spectrophotometric constants for common hemoglobin derivatives in human, dog and rabbit blood. J Biol Chem 1932;98:719-33.

Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med1963;61:882-8.

Constandinou C, Henderson N, Iredale JP. Modeling liver fibrosis in rodents. Method Mol Med 2005;117:237-50.

De Andrade Belo MA, Soares VE, De Souza LM, et al. Hepatoprotective treatment attenuates oxidative damages induced by carbon tetrachloride in rats. Exp Toxicol Pathol 2012;64(3):155-65.

Yamaoka K, Kataoka T, Nomura T, et al. Inhibitory effects of prior low-dose X-ray irradiation on carbon tetrachloride-induced hepatopathy in acatalasemic mice. J Radiat Res 2004;45(1):89-95.

Chen W, Kennedy D, Kojima A, et al. Polyamines and thiols in the cytoprotective effect of L-cysteine and Lmethionine on carbon tetrachloride-induced hepatotoxicity. Amino Acids 2000;18(4):319-27.

Halliwell B, Gutteridge J. Oxygen free radicals and iron in relation to biology and medicine: some problems and concepts. Arch Biochem Biophys 1986;246(2):501-14.

Dalle-Donne I, Rossi R, Giustarini D, e al. Protein carbonyl groups as biomarkers of oxidative stress. ClinChim Acta 2003;329(1-2):23-38.

Geetha A, Lakshmi Priya MD, Jeyachristy SA, et al. Level of oxidative stress in the red bloodcells of patients with liver cirrhosis. Indian J Med Res 2007;126(3):204.-10

Rockey DC, Chung JJ. Reduced nitric oxide production by endothelial cells in cirrhotic rat liver: endothelial dysfunction in portal hypertension. Gastroenterology 1998;114(2):344-51.

Zimmermann H, Kurzen P, Klossner W, et al. Decreased constitutive hepatic nitric oxide synthase expression in secondary biliary fibrosis and its changes after Roux-en-Y choledocho-jejunostomy in the rat. J Hepatol 1996;25(4):567-73.

Magnani M, Serafini G, Stocchi V, et al. Solubilization, purification, and properties of rabbit brain hexokinase. Arch Biochem Biophys 1982;216(2):449-54.


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