Abstract
Nonalcoholic steatohepatitis (NAFLD) is a progressive form of liver disease that leads to advanced fibrosis. The present study was designed to assess the hepatoprotective effect of thymoquinone (TQ) on liver functions, insulin resistance, and PPAR-γ expression in NAFLD. Rats were divided into two main groups: one fed with normal rat chow diet and the other with high-fat high-cholesterol diet group for 6 weeks. Every group was subdivided into three subgroups (n = 8): treated with saline, low dose TQ (10 mg/kg), high dose TQ (20 mg/kg). High fat high cholesterol diet caused marked liver damage as noted in histopathology and significant increase in liver index, liver enzymes. There was significant increase in the insulin resistance, serum cholesterol, triglyceride, PPAR-γ gene overexpression with significant decrease in HDL. Additionally, oxidative stress increased by measuring MDA associated with significant decrease in serum total antioxidant capacity. As markers of inflammation, hepatic TNF-α was significantly increased with decrease in IL10. Further, there was increase in BAX protein with decrease in Bcl as compared to control group. This model of 6 weeks high-fat high-cholesterol diet showed minimal fibrosis as noticed by increase MMP2 and Masson trichrome satin. Co-treatment with TQ improved all previous parameters. High dose was more effective, although mostly non-statistically significant. TQ may have a promising agent to improve hepatic steatosis, oxidative stress; inflammatory, apoptotic status, fibrosis and so prevent liver damage in patients with NAFLD. Although PPAR-γ was significantly under-expressed by TQ, insulin resistance was improved significantly suggesting a role of liver damage.
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Abdel-Wahab WM (2014) Thymoquinone attenuates toxicity and oxidative stress induced by bisphenol A in liver of male rats. Pak J Biol Sci 17(11):1152–1160
Abukhader MM (2012) The effect of route of administration in thymoquinone toxicity in male and female rats. Indian J Pharm Sci 74(3):195–200
Ahmad A, Husain A, Mujeeb M, Khan SA, Najmi AK, Siddique NA, Damanhouri ZA, Anwar F (2013) A review on therapeutic potential of Nigella sativa: a miracle herb. Asian Pac J Trop Biomed 3(5):337–352
Ahmad S, Beg ZH (2013) Elucidation of mechanisms of actions of thymoquinone-enriched methanolic and volatile oil extracts from Nigella sativa against cardiovascular risk parameters in experimental hyperlipidemia. Lipids Health Dis 12:86
Asgary S, Sahebkar A, Goli-Malekabadi N (2015) Ameliorative effects of Nigella sativa on dyslipidemia. J Endocrinol Invest 38(10):1039–1046
Basaranoglu M, Basaranoglu G, Senturk H (2013) From fatty liver to fibrosis: a tale of “second hit”. World J Gastroenterol 19(8):1158–1165
Bataller R, Brenner DA (2005) Liver fibrosis. J Clin Invest 115(2):209–218
Bechmann LP, Kocabayoglu P, Sowa JP, Sydor S, Best J, Schlattjan M, Beilfuss A, Schmitt J, Hannivoort RA, Kilicarslan A, Rust C, Berr F, Tschopp O, Gerken G, Friedman SL, Geier A, Canbay A (2013) Free fatty acids repress small heterodimer partner (SHP) activation and adiponectin counteracts bile acid-induced liver injury in superobese patients with nonalcoholic steatohepatitis. Hepatology 57(4):1394–1406
Bedoucha M, Atzpodien E, Boelsterli UA (2001) Diabetic KKAy mice exhibit increased hepatic PPARgamma1 gene expression and develop hepatic steatosis upon chronic treatment with antidiabetic thiazolidinediones. J Hepatol 35(1):17–23
Brunt EM, Tiniakos DG (2010) Histopathology of nonalcoholic fatty liver disease. World J Gastroenterol 16(42):5286–5296
Burits M, Bucar F (2000) Antioxidant activity of Nigella sativa essential oil. Phytother Res 14(5):323–328
Day CP, James OF (1998) Steatohepatitis: a tale of two “hits”? Gastroenterology 114(4):842–845
Dowman JK, Tomlinson JW, Newsome PN (2010) Pathogenesis of non-alcoholic fatty liver disease. QJM 103(2):71–83
El-Sheikh AA, Morsy MA, Abdalla AM, Hamouda AH, Alhaider IA (2015) Mechanisms of Thymoquinone Hepatorenal Protection in Methotrexate-Induced Toxicity in Rats. Mediat Inflamm 2015:859383
Elinav E, Pappo O, Sklair-Levy M, Margalit M, Shibolet O, Gomori M, Alper R, Thalenfeld B, Engelhardt D, Rabbani E, Ilan Y (2006) Amelioration of non-alcoholic steatohepatitis and glucose intolerance in ob/ob mice by oral immune regulation towards liver-extracted proteins is associated with elevated intrahepatic NKT lymphocytes and serum IL-10 levels. J Pathol 208(1):74–81
Federico A, Niosi M, Vecchio Blanco CD, Loguercio C (2008) Emerging drugs for non-alcoholic fatty liver disease. Expert Opin Emerg Drugs 13(1):145–158
Friedman SL (2004) Mechanisms of disease: mechanisms of hepatic fibrosis and therapeutic implications. Nat Clin Pract Gastroenterol Hepatol 1(2):98–105
Gavrilova O, Haluzik M, Matsusue K, Cutson JJ, Johnson L, Dietz KR, Nicol CJ, Vinson C, Gonzalez FJ, Reitman ML (2003) Liver peroxisome proliferator-activated receptor gamma contributes to hepatic steatosis, triglyceride clearance, and regulation of body fat mass. J Biol Chem 278(36):34268–34276
Grattagliano I, Vendemiale G, Caraceni P, Domenicali M, Nardo B, Cavallari A, Trevisani F, Bernardi M, Altomare E (2000) Starvation impairs antioxidant defense in fatty livers of rats fed a choline-deficient diet. J Nutr 130(9):2131–2136
Hevener AL, He W, Barak Y, Le J, Bandyopadhyay G, Olson P, Wilkes J, Evans RM, Olefsky J (2003) Muscle-specific Pparg deletion causes insulin resistance. Nat Med 9(12):1491–1497
Hickman IJ, Clouston AD, Macdonald GA, Purdie DM, Prins JB, Ash S, Jonsson JR, Powell EE (2002) Effect of weight reduction on liver histology and biochemistry in patients with chronic hepatitis C. Gut 51(1):89–94
Hosseini M, Zakeri S, Khoshdast S, Yousefian FT, Rastegar M, Vafaee F, Kahdouee S, Ghorbani F, Rakhshandeh H, Kazemi SA (2012) The effects of Nigella sativa hydro-alcoholic extract and thymoquinone on lipopolysaccharide-induced depression like behavior in rats. J Pharm Bioallied Sci 4(3):219–225
Houghton PJ, Zarka R, de las Heras B, Hoult JR (1995) Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med 61(1):33–36
Jaswal A, Sinha N, Bhadauria M, Shrivastava S, Shukla S (2013) Therapeutic potential of thymoquinone against anti-tuberculosis drugs induced liver damage. Environ Toxicol Pharmacol 36(3):779–786
Jeong YY, Lee YL, Lee KM, Jong-Yeon JK (2009) The Effects of oenanthe javanica extracts on hepatic fat accumulation and plasma biochemical profiles in a non-alcoholic fatty liver disease model. J Korean Soc Appl Biol Chem 52(6):632–637
Kanter M (2009) Protective effects of thymoquinone on streptozotocin-induced diabetic nephropathy. J Mol Histol 40(2):107–115
Kersten S, Desvergne B, Wahli W (2000) Roles of PPARs in health and disease. Nature 405(6785):421–424
Kim MH, Kang KS (2012) Isoflavones as a smart curer for non-alcoholic fatty liver disease and pathological adiposity via ChREBP and Wnt signaling. Prev Med 54(Suppl):S57–S63
Koracevic D, Koracevic G, Djordjevic V, Andrejevic S, Cosic V (2001) Method for the measurement of antioxidant activity in human fluids. J Clin Pathol 54(5):356–361
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods 25(4):402–408
Mahady SE, George J (2012) Management of nonalcoholic steatohepatitis: an evidence-based approach. Clin Liver Dis 16(3):631–645
Malhi H, Bronk SF, Werneburg NW, Gores GJ (2006) Free fatty acids induce JNK-dependent hepatocyte lipoapoptosis. J Biol Chem 281(17):12093–12101
Mansour MA, Ginawi OT, El-Hadiyah T, El-Khatib AS, Al-Shabanah OA, Al-Sawaf HA (2001) Effects of volatile oil constituents of Nigella sativa on carbon tetrachloride-induced hepatotoxicity in mice: evidence for antioxidant effects of thymoquinone. Res Commun Mol Pathol Pharmacol 110(3–4):239–251
Marra F, Lotersztajn S (2013) Pathophysiology of NASH: perspectives for a targeted treatment. Curr Pharm Des 19(29):5250–5269
Memon RA, Tecott LH, Nonogaki K, Beigneux A, Moser AH, Grunfeld C, Feingold KR (2000) Up-regulation of peroxisome proliferator-activated receptors (PPAR-alpha) and PPAR-gamma messenger ribonucleic acid expression in the liver in murine obesity: troglitazone induces expression of PPAR-gamma-responsive adipose tissue-specific genes in the liver of obese diabetic mice. Endocrinology 141(11):4021–4031
Mihara M, Uchiyama M (1978) Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 86(1):271–278
Moschen AR, Wieser V, Tilg H (2012) Adiponectin: key player in the adipose tissue-liver crosstalk. Curr Med Chem 19(32):5467–5473
Nagi MN, Mansour MA (2000) Protective effect of thymoquinone against doxorubicin-induced cardiotoxicity in rats: a possible mechanism of protection. Pharmacol Res 41(3):283–289
Norris AW, Chen L, Fisher SJ, Szanto I, Ristow M, Jozsi AC, Hirshman MF, Rosen ED, Goodyear LJ, Gonzalez FJ, Spiegelman BM, Kahn CR (2003) Muscle-specific PPARgamma-deficient mice develop increased adiposity and insulin resistance but respond to thiazolidinediones. J Clin Invest 112(4):608–618
Ojha S, Azimullah S, Mohanraj R, Sharma C, Yasin J, Arya DS, Adem A (2015) Thymoquinone Protects against Myocardial Ischemic Injury by Mitigating Oxidative Stress and Inflammation. Evid Based Complement Alternat Med 2015:143629
Oliveira CP, da Costa Gayotto LC, Tatai C, Della Bina BI, Janiszewski M, Lima ES, Abdalla DS, Lopasso FP, Laurindo FR, Laudanna AA (2002) Oxidative stress in the pathogenesis of nonalcoholic fatty liver disease, in rats fed with a choline-deficient diet. J Cell Mol Med 6(3):399–406
Pan MH, Lai CS, Tsai ML, Ho CT (2014) Chemoprevention of nonalcoholic fatty liver disease by dietary natural compounds. Mol Nutr Food Res 58(1):147–171
Pari L, Sankaranarayanan C (2009) Beneficial effects of thymoquinone on hepatic key enzymes in streptozotocin-nicotinamide induced diabetic rats. Life Sci 85(23–26):830–834
Peng XD, Dai LL, Huang CQ, He CM, Yang B, Chen LJ (2009) Relationship between anti-fibrotic effect of Panax notoginseng saponins and serum cytokines in rat hepatic fibrosis. Biochem Biophys Res Commun 388(1):31–34
Ratziu V, Charlotte F, Bernhardt C, Giral P, Halbron M, Lenaour G, Hartmann-Heurtier A, Bruckert E, Poynard T (2010) Long-term efficacy of rosiglitazone in nonalcoholic steatohepatitis: results of the fatty liver improvement by rosiglitazone therapy (FLIRT 2) extension trial. Hepatology 51(2):445–453
Rockey DC, Boyles JK, Gabbiani G, Friedman SL (1992) Rat hepatic lipocytes express smooth muscle actin upon activation in vivo and in culture. J Submicrosc Cytol Pathol 24(2):193–203
Satapathy SK, Garg S, Chauhan R, Sakhuja P, Malhotra V, Sharma BC, Sarin SK (2004) Beneficial effects of tumor necrosis factor-alpha inhibition by pentoxifylline on clinical, biochemical, and metabolic parameters of patients with nonalcoholic steatohepatitis. Am J Gastroenterol 99(10):1946–1952
Schwenger KJ, Allard JP (2014) Clinical approaches to non-alcoholic fatty liver disease. World J Gastroenterol 20(7):1712–1723
Sugii S, Olson P, Sears DD, Saberi M, Atkins AR, Barish GD, Hong SH, Castro GL, Yin YQ, Nelson MC, Hsiao G, Greaves DR, Downes M, Yu RT, Olefsky JM, Evans RM (2009) PPARgamma activation in adipocytes is sufficient for systemic insulin sensitization. Proc Natl Acad Sci U S A 106(52):22504–22509
Takahashi Y, Soejima Y, Fukusato T (2012) Animal models of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World J Gastroenterol 18(19):2300–2308
Tessari P, Coracina A, Cosma A, Tiengo A (2009) Hepatic lipid metabolism and non-alcoholic fatty liver disease. Nutr Metab Cardiovasc Dis 19(4):291–302
Tilg H, Moschen AR (2010) Evolution of inflammation in nonalcoholic fatty liver disease: the multiple parallel hits hypothesis. Hepatology 52(5):1836–1846
Tontonoz P, Hu E, Spiegelman BM (1994) Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell 79(7):1147–1156
Umar S, Hedaya O, Singh AK, Ahmed S (2015) Thymoquinone inhibits TNF-alpha-induced inflammation and cell adhesion in rheumatoid arthritis synovial fibroblasts by ASK1 regulation. Toxicol Appl Pharmacol 287(3):299–305
Utzschneider KM, Kahn SE (2006) Review: The role of insulin resistance in nonalcoholic fatty liver disease. J Clin Endocrinol Metab 91(12):4753–4761
Wieckowska A, Papouchado BG, Li Z, Lopez R, Zein NN, Feldstein AE (2008) Increased hepatic and circulating interleukin-6 levels in human nonalcoholic steatohepatitis. Am J Gastroenterol 103(6):1372–1379
Williams CD, Stengel J, Asike MI, Torres DM, Shaw J, Contreras M, Landt CL, Harrison SA (2011) Prevalence of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis among a largely middle-aged population utilizing ultrasound and liver biopsy: a prospective study. Gastroenterology 140(1):124–131
Wree A, Kahraman A, Gerken G, Canbay A (2011) Obesity affects the liver—the link between adipocytes and hepatocytes. Digestion 83(1–2):124–133
Xu ZJ, Fan JG, Ding XD, Qiao L, Wang GL (2010) Characterization of high-fat, diet-induced, non-alcoholic steatohepatitis with fibrosis in rats. Dig Dis Sci 55(4):931–940
Yilmaz B, Sahin K, Bilen H, Bahcecioglu IH, Bilir B, Ashraf S, Halazun KJ, Kucuk O (2015) Carotenoids and non-alcoholic fatty liver disease. Hepatobiliary Surg Nutr 4(3):161–171
Zheng H, Li S, Ma L, Cheng L, Deng C, Chen Z, Xie C, Xiang M, Jiang W, Chen L (2011) A novel agonist of PPAR-gamma based on barbituric acid alleviates the development of non-alcoholic fatty liver disease by regulating adipocytokine expression and preventing insulin resistance. Eur J Pharmacol 659(2–3):244–251
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Awad, A.S.M., Abd Al Haleem, E.N., El-Bakly, W.M. et al. Thymoquinone alleviates nonalcoholic fatty liver disease in rats via suppression of oxidative stress, inflammation, apoptosis. Naunyn-Schmiedeberg's Arch Pharmacol 389, 381–391 (2016). https://doi.org/10.1007/s00210-015-1207-1
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DOI: https://doi.org/10.1007/s00210-015-1207-1