Ameliorative Potential of Thiazolidinediones and Statins on Experimentally Induced Non-Alcoholic Steatohepatitis

Ahmed M Kabel^{1, 2,} and Hany M Borg³

¹Department of Clinical Pharmacy, College of Pharmacy, Taif University, Taif, Saudi Arabia

²Department of Pharmacology, Faculty of Medicine, Tanta University, Tanta, Egypt

³Department of Physiology, Faculty of Medicine, Kafrelsheikh University, Egypt

Cite this paper:
Ahmed M Kabel and Hany M Borg. Ameliorative Potential of Thiazolidinediones and Statins on Experimentally Induced Non-Alcoholic Steatohepatitis. World Journal of Nutrition and Health. 2014; 2(4):52-57. doi: 10.12691/jnh-2-4-2

Abstract

Non-alcoholic steatohepatitis (NASH) is a chronic liver disease that may progress to advanced fibrosis and cirrhosis. Yet, its pathogenesis is not fully understood. The aim of this work was to study the possible protective effect of atorvastatin and pioglitazone on non-alcoholic steatohepatitis induced by high fat diet (HFD) in rats. Sixty male albino rats were divided into two main groups, the normal control group (ten rats) and HFD group (fifty rats) which was further subdivided into five subgroups (ten rats each) one of them received HFD+normal saline (NASH group), the 2nd received HFD + vehicle (gum tragacanth), the 3rd received HFD+atorvastatin, the 4th received HFD+ pioglitazone and the last group received HFD+atorvastatin and pioglitazone. At the end of the study, animals were killed and blood samples were collected for estimations of serum ALT, AST, ALP, TNF-α, triglycerides, total cholesterol, glucose, insulin and HOMA-IR. Also, samples of liver were taken and studied for hepatic triglycerides, malondialdehyde, free fatty acids and histopathological analysis. Atorvastatin lowered efficiently serum ALT, AST, ALP, TNF-α, total cholesterol, triglycerides, hepatic triglycerides, malondialdehyde and free fatty acids, together with marked improvement in histopathology of liver steatohepatitis, but, produced insignificant effects on fasting blood glucose, serum insulin and HOMA-IR. On the other hand, administration of pioglitazone, whether alone or in combination with atorvastatin induced significant improvement of all the above mentioned parameters. In conclusion, if the prominent feature in NASH is insulin resistance, we recommend the use of pioglitazone, while atorvastatin will be needed if the prominent features is hyperlipidemia and both drugs simultaneously if there are both hyperlipidemia and insulin resistance.

Keywords:
statins thiazolidinediones non-alcoholic steatohepatitis rats

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References:

[1]	Adams LA and Angulo P (2006). Treatment of non-alcoholic fatty liver disease. Postgraduate Medical J; 82: 315-322.
[2]	Angulo P, Keach JC, Batts KP, Lindor KD (1999). Independent predictors of liver fibrosis in patients with nonalcoholic steatohepatitis. Hepatology; 30: 1356-1362.
[3]	Blighard EG and Dyer MG (1995). A rapid method of total lipid extractions and purification. Cam J Biochem Physiol; 27 (8): 911-7.
[4]	Boden G (1997). Role of fatty acids in the pathogenesis of insulin resistance and NIDDM. Diabetes; 46: 3-10.
[5]	Bonora E, Targer G, Alberiche M, Bonadonna RC, Saggiani F, Zenere MB, Monauni T, Muggeo M (2000). Homeostasis model assessment closely mirrors the glucose clamp technique in the assessment of insulin sensitivity: studies in subjects with various degrees of glucose tolerance and insulin sensitivity. Diabetes Care; 23 (1): 57-63.
[6]	Browning JD and Horton JD (2004). Molecular mediators of hepatic steatosis and liver injury. J Clin Invest; 114: 147-152.
[7]	Bugianesi E, Manzini P, D antico S, Vanni E, Longo F, Leone N, Massarenti P, Piga A, Marchesini G, Rizzetto M (2004). Relative contribution of iron burden, HFE mutations and insulin resistance to fibrosis in non-alcoholic fatty liver. Hepatology; 39: 179: 187.
[8]	Chitturi S and Farrell GC (2001). Etiopathogenesis of non-alcoholic steatohepatitis. Semin Liver Dis; 21: 27-41.
[9]	Cohen B, Noick D, Rubinstein M (1996). Modulation of insulin activities by leptin. Science; 274: 1185-1188.
[10]	Comar KM, Sterling RK (2006). Reviewarticledrug therapy for non-alcoholic fatty liver disease. Aliment Pharmacol Ther; 23: 207-15.
[11]	Da Silva Morais A, Lebrun V, Abarca-Quinones J, Brichard S, Hue L, Guigas B, Viollet B, Leclercq IA (2009). Prevention of steatohepatitis by pioglitazone: implication of adiponectin-dependent inhibition of SREBP-1c and inflammation Hepatol; 50 (3): 489-500.
[12]	Day CP and James O (1998). Steatohepatitis: Atale of two hits? Gastroentrology; 114: 842-845.
[13]	Fan JG, Zhong L, Xu ZJ, Tia LY, Ding XD, Li MS, Wang GL (2003). Effects of low calory diet on seteatohepatitis in rats with obesity and hyperlipidemia. World J Gastroentrol; 9 (9): 2045-2049.
[14]	Fossati P and Prencipe I (1980). Serum triglycerides dtermined colorimercially with an enzyme that produces hydrogen peroxide. Clin Chem; 28: 2077.
[15]	Fruebis J, Tsao TS, Javorschi S, Ebbets-Reed D, Erickson MR, Yen FT, et al. (2001). Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice. Proc Natl Acad Sci USA; 98: 2005-2010.
[16]	Gianluca SP, Cinzia C, Stefania S, Giabnna F, Tiziana B, Marco M, Samuele DM, Liliana N, Renata S, Alessia O, Deborah B, Soren S, Antonio B, Alessndro C (2006). A model of insulin resistance and nonalcoholic steatohepatitis in rats: Role of peroxisome proliferators activated receptor-α and n-3 polyunsaturated fatty acid treatment on liver injury. Am J Pathol.; 169 (3): 846-860.
[17]	Gomez-Dominguez E, Gisbert JP, Moreno-Monteagudo JA et al., (2006). A pilot study of atorvastatin treatment in dyslipidemia, nonalcoholic fatty liver patients. Aliment Pharmacol Ther; 23: 1643-7.
[18]	Grip O, Janciauskiene S, Lindgren S (2002). Atorvastatin activates PPAR gamma and attenuates the inflammatory response in human monocytes. Inflamm Res; 51: 58-62.
[19]	Hotamisligil GS, Peraldi SP, Budaari A, Ellis R, White MF, Speegelman BM (1996). IRS-1 mediated inhibition of insulin receptor tyrosine kinase activity in TNF-α and obesity induced insulin resistance. Science; 271: 665-668.
[20]	Hyogo H, Tazuma S, Arihiro K, Iwamoto K, Nabeshima Y, Inoue M, Ishitobi T, Nonaka M, Chayama K (2008). Efficacy of atorvastatin for the treatment of nonalcoholic steatohepatitis with dyslipidemia. Metabolism; 57 (12): 1711-8.
[21]	Imeryuz N, Tahan V, Sonsuz A et al. (2007). Iron preloading aggravates nutritional steatohepatitis in rats by increasing apoptotic cell death. J Hepatol; 47: 851-859.
[22]	Ishikawa E, Hashida S, Taanaka K, Kohno T (1989). Development and application of ultra-sensitive enzyme immunoassays for antigens and antibodies. Clin Chem Acts; 185: 223-30.
[23]	Itaya K and Kadowaki T (1969). An improved method for colorimetric determination of free fatty acids. Clin Chim Acta; 26: 401.
[24]	Kabel AM, Abdel-Rahman MN, El-Sisi Ael-D, Haleem MS, Ezzat NM, El Rashidy MA (2013). Effect of atorvastatin and methotrexate on solid Ehrlich tumor. Eur J Pharmacol; 713 (1-3): 47-53.
[25]	Kallwitz ER, Mclachlan A, Cotler S (2008). Role of peroxisome prolifeator-activated receptors in the pathogenesis and treatment of nonalcoholic fatty liver disease. World J Gastroenterol; 14: 22-28.
[26]	Kamada Y, Tamura S, Kiso S, Matsumoto H, Saji Y, Yoshida Y, et al (2003). Enhanced carbon tetrachloride-induced liver fibrosis in mice lacking adiponectin. Gastroenterology; 125: 1796-1807.
[27]	Kawaguchi K, Sakaida I, Tsuchiya M, Omori K, Takami T, Okita K (2004). Pioglitazone prevents hepatic steatosis, fibrosis and-altered lesion in rat liver cirrhosis induced by a choline-deficient L-amino acid-defined diet. Biochem and Biophys Res Comm; 315:187-195.
[28]	Koruk M, Taysi S, Saas MC, Yilmaz O, Akcay F, Karakok M (2004). Oxidative stress and enzymatic antioxidant status in patients withy non-alcoholic steatohepatitis. Ann Cli Lab Sci Winter; 34 (1): 57-62.
[29]	Leclercq IA, Farrell GC, Sempoux C, Dela Pena A, Hormans Y (2004). Curcumin inhibits NF-kappa B activation and reduces the severity of experimental steatohepatitis in mice. J Hepatol; 41: 926-934.
[30]	Lieber CS, Leo MA, Mak KM, Xu Y, Cao Q, Ren C, Ponomarenko A, DeCarli LM (2004). Model of nonalcoholic steatohepatitis. Am J Clin Nutr; 79 (3): 350-351.
[31]	Lowry OH, Rosebrough N, Farr AL et al. (1951). Protein measurement with folin phenol reagent. J Biol Chem; 193: 265-275.
[32]	Maddux BA, Sbraccia P, Kumakura (1995). Membrane glycoprotein PC1 and insulin resistance in non insulin dependent diabetes mellitus. Nature; 373: 448-451.
[33]	Metha K, Vanthiel DH, Shah N et al.,(2002). Nonalcoholic fatty liver disease; pathogenesis and the role of antioxidants. Nutr Rev; 60: 289-293.
[34]	Miyazaki Y, Mahankali A, Wajcberg E, Bajaj M, Mandarino LJ, DeFronzo RA (2004). Effect of pioglitazone on circulating adipocytokine levels and insulin sensitivity in type 2 diabetic patients. J Clin endocrinol Metab; 89: 4312-4319.
[35]	Nawrocki AR, Rajala MW, Tomas E, Pajvani UB, Saha AK, Trumbauer ME, et al., (2006). Mice lacking adiponectin show decreased hepatic insulin sensitivity and reduced responsiveness to peroxisome proliferator-activated receptor gamma agonists. J Biol Chem; 281: 2654-2660.
[36]	Ohkawa H, Ohishi N and Yagi K (1979). Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem; 95: 351-358.
[37]	Orasanu G, Ziouzenkova O, Devchand PR, Nehra V, Hamdy O,Horton ES, et al. (2008). The peroxisome proliferator-activated receptorgamma agonist pioglitazone represses inflammation in a peroxisome proliferator-activated receptor-alpha-dependent manner in vitro and in vivo in mice. J Am Coll Cardiol; 52: 869-881.
[38]	Paradise V, Perlemuter G, Bonovoust F, et al., (2001). High glucose and hyperinsulinemia stimulate connective tissue growth factor expression: a potential mechanism involved in progression to fibrosis in non-alcholic steatohepatitis. Hepatology; 34: 738-44.
[39]	Ping X, Zhang X, Li Y, Yu C, Xu L, Xu GY (2006). Research on the protection effect of pioglitazone for non-alcoholic fatty liver isease (NAFLD) in rats. J Zhejiang Uni Sci B; 7(8): 627-633.
[40]	Reitman S and Frankel S (1957). A colorimetric method for determination of serum glutamic oxaloacetic and glutamic pyruvic transaminase. Am J Clin Pathol; 28: 56-63.
[41]	Richmoud W (1973): Preparations and properties of a cholesterol oxidase from Nocardia sp. And its application to the enzymatic assay of total cholesterol in serum. Chin Chem; 19 (12): 1350-6
[42]	Robertson G, Leclercq I, Farrell GC (2001). Nonalcoholic steatosis and steatohepatitis. II. Cytochrome P-450 enzymes and oxidative stress. Am J Physiol: Gastrointest Liver Physiol; 281: 1135-1139.
[43]	Sanyal AJ, Campbell-Sargent C, Mirshahi F, Rizzo WB, Contos MJ, Sterling RK, Luketic VA, Shiffman MKL, Clore JN (2001). Non-alcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalities. Gastroenterol; 120: 1183-1192.
[44]	Seki S, Kitada T, Sakaguchi H (2005). Clincopathological significance of oxidative cellular damage in non-alcoholic fatty liver diseases. Hepatol Res; 33 (2): 132-4.
[45]	Song XY, Gu M, Jin WW, Klinman DM, Wahl SM (1998). Plasmid DNA encoding transforming growth factor-beta1 supresses chronic disease in a streptococal cell wall induced arthritis model. Jclin Invest; 101 (12) 2615:21.
[46]	Stepaniake JA, Gould KE, Sun D, Swanborg RH (1995). A comparative study of experimental antiimmune encephalomyelitis in Lewis and DA rats. J Immunol; 155(5): 2762-9.
[47]	Tahan V, Eren F, Avsar E et al. (2007). Rosiglitazone attenuates liver inflammation in a rat model of nonalcoholic steatohepatitis. Dig Dis Sci; 52: 3465-3472.
[48]	Takaki A, Kawai D, Yamamoto K (2014). Molecular Mechanisms and New Treatment Strategies for non-Alcoholic Steatohepatitis (NASH). Int J Mol Sci; 15: 7352-7379.
[49]	Tamura Y, Kawamori R (2006). Insulin resistance. Nippon Rhinsho; 64(6):1071-4.
[50]	Teranishi T, Ohara T, Maeda K, Zenibayashi M, Kouyama K, Hirota Y, et al. (2007). Effects of pioglitazone and metformin on intracellular lipid content in liver and skeletal muscle of individuals with type 2 diabetes mellitus. Metabolism; 56:1418-1424.
[51]	Torres DM and Harrison SA (2008). Steatohepatitis: a tale of two hits? Gastroentrology; 114: 842-845.
[52]	Trinder P (1969). Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor. Ann Clin Biochem; 6: 24-25.
[53]	Von Knethen A and Brune B (2003). PPAR gamma-an important regulator of monocyte/macrophage function. Arch Immunol Ther Exp (Warsz); 51: 219-226.
[54]	Xu A, Wang Y, Keshaw H, Xu LY, Lam KS, Cooper GJ (2003). The fat derived hormone adiponectin alleviates alcoholic and nonalcoholic fatty liver diseases in mice. J Clin Invest; 112: 91-100.
[55]	Xu P, Zhang X, Li Y, Yu L, Xu G (2006). Research on the protection of pioglitazone for non-alcoholic fatty liver disease (NAFDL) in rats. Journal of Zhejiang university Science B; 7 (8): 627-633.
[56]	Yokohama S, Yomeda M, Hameda M et al., (2004). Theraputic efficacy of an angiotensin II receptors antagonist in patients with non-alcoholic steatohepatitis. Hepatology; 40: 1222-5.
[57]	Yokota T, Oritani K, Takahashi I, Ishikawa J, Matsuyama A, Ouchi N, et al (2000). Adiponectin, a new member of the family of soluble defense collagens, negatively regulates the growth of myelomonocytic progenitors and the functions of macrophages. Blood; 96: 1723-1732.