- Email: [email protected]
1258 ORAL BIFIDOBACTERIUM ADOLESCENTIS SUPPLEMENTATION ATTENUATES NONALCOHOLIC STEATOHEPATITIS (NASH) IN A MOUSE MODEL
POSTERS experiments suggest that PNPLA3 expression depends on the types of dietary fatty acids with polyunsaturated fatty acids inducing and monounsaturated fatty acids inhibiting PNPLA3 mRNA. 1256 DIETARY SUPPLEMENTATION WITH OLIVE OIL WITH OR WITHOUT N-3 PUFA DIFFERENTIALLY AFFECTS INFLAMMATION AND PROGRESSION OF EXPERIMENTAL NONALCOHOLIC STEATOHEPATITIS A. Provenzano1 , S. Milani1 , F. Vizzutti1 , W. Delogu1 , N. Navari1 , E. Novo2 , G. Laffi1 , M. Parola2 , F. Marra1 . 1 University of Florence, Florence, 2 University of Turin, Turin, Italy E-mail: [email protected] Background/aims: While it is established that excessive calorie intake plays a role in the pathogenesis of nonalcoholic steatohepatitis, the significance of individual dietary factors is still elusive. Supplementation with n-3 polyunsaturated fatty acids (PUFA) has been shown to ameliorate fatty liver in experimental models, and clinical studies are underway. In this study we compared the effects of supplementation with olive oil (mostly composed of oleic acid, 18:1 n-9) or olive oil and n-3 PUFA on the progression of experimental steatohepatitis. Methods: Balb/C mice (≥5 mice/group) were fed a methionine and choline deficient (MCD) diet or a control diet for 4 or 8 weeks. At the same time, mice were supplemented with n-3 PUFA (eicosapentaenoic and docosahexahenoic acid, 25 mg together with 75 mg olive oil), or olive oil alone (OO, 100 mg), two times a week by intragastric gavage Results: Aminotransferase levels were not different comparing mice supplemented with n-3 or OO after 4w of MCD diet, while after 8w mice on MCD/n-3 had more severe necroinflammation compared to MCD/OO (ALT: 373±170 vs. 87±29 UI/L, P < 0.05). Liver/body weight ratio was significantly lower in MCD/n-3 mice after 8w of treatment (5.1±0.8 vs. 6.8±1.0%, P < 0.05). At 8w, the score of inflammation and fibrosis was significantly higher in mice receiving MCD/n-3 than in MCD/OO animals, with a marked increase in the number of lipogranulomas (26.4±8.4 vs. 5.1±5. per field, P < 0.001). A significant increase in intrahepatic expression of TNF-alpha and CCL2 was observed at both 4w and 8w in MCD/n-3 mice, which also had increased expression of CD11b at 4 weeks. In addition, increased transcript levels of the profibrogenic genes TIMP-1 and TGF-beta, and lower expression of PPAR-alpha was observed in MCD/n-3 mice. After 8 week of MCD diet, portal pressure was higher in mice receiving n-3 than in those on OO (5.1±1.4 vs. 7.0±0.9 mmHg, P < 0.05). No major differences were observed comparing n-3 and OO supplementation together with a control diet. Conclusions: In a model of steatohepatiits, supplementation with n-3 PUFA and OO is associated with more severe necroinflammation and fibrosis than in mice treated with OO only. 1257 SORTILIN KNOCKOUT MICE DISPLAY ATTENUATED HEPATIC INJURY AND INFLAMMATION IN METABOLIC STRESS MODELS L. Rabinowich1 , R. Amir2 , E. Hubel2 , Z. Halpern2 , E.M. Santo2 , S. Fishman2 , I. Zvibel2 . 1 Internal Medicine D, 2 Gastroenterology, Tel Aviv Sourasky Medical Center, Tel Aviv University, Tel Aviv, Israel E-mail: [email protected] Background and Aims: Sortilin is a member of vacuolar protein sorting 10 protein-domain receptors, whose role is to direct trafficking of newly synthesized molecules from the trans-Golgi network to regulated secretion pathways or to endosomes and lysosomes. Recently, genome wide studies have pinpointed to sortilin in hepatocytes as a key molecule regulating serum LDL. In adipocytes and muscle, sortilin regulates transfer of glut4 from
intracellular vesicles to the cell surface, thus mediating insulindependent glucose uptake. Based on its several metabolic roles, we hypothesized that sortilin affects whole body glucose metabolism and hepatic steatosis and inflammation. With respect to hepatic fat accumulation, it has been shown that increased lipogenesis which ends up with triglyceride accumulation serves as a favorable fat store protecting the liver from inflammation. Methods: C57BL/6 (WT) and sortilin −/− male mice were fed regular chow (RC) and Western diet for two months and methioninecholine deficient diet (MCD) for 5 weeks. Results: Sortilin−/− mice have significantly reduced body weight and visceral fat both after RC and Western diet, despite similar food intake. In addition, sortilin−/− mice have significantly lower serum cholesterol and exhibit better glucose tolerance tests. Interestingly, hepatic lipogenesis was increased, as demonstrated by increased mRNA expression of fatty acid synthase and stearoyl CoA desaturase. Hepatic expression of phosphorylated acetyl CoA carboxylase was significantly lower in sortilin−/− mice, also indicative of increased lipogenesis. In addition, livers of sortilin −/− mice showed reduced mRNA levels of hepatic inflammatory, insulin resistance-inducing cytokines TNFa and PAI-1. Adipose tissue of sortilin−/− displays significantly smaller adipocyte size and decreased mRNA levels of inflammatory cytokines TNFa, PAI1and MCP-1. Muscle tissue of these mice express higher levels of glut4 and differentiation marker myogenin. These data suggest that sortilin −/− have better metabolic parameters and attenuated liver inflammation, even though they exhibit increased hepatic lipogenesis. In accordance with the above, sortilin−/− mice fed MCD show attenuated liver damage as expressed by 5-fold lower AST (143 vs 875unit/L) and twice lower ALT (132 vs 282unit/L), yet stronger steatosis. Conclusion: Lack of sortilin induces a beneficial metabolic phenotype and attenuated liver inflammation in metabolic stress models, accompanied by increased lipogenesis. 1258 ORAL BIFIDOBACTERIUM ADOLESCENTIS SUPPLEMENTATION ATTENUATES NONALCOHOLIC STEATOHEPATITIS (NASH) IN A MOUSE MODEL A. Reichold, S. Brenner, A. Spruss, A. Rings, K. Forster-Fromme, ¨ I. Bergheim, S.C. Bischoff. Department of Nutritional Medicine (180a), University of Hohenheim, Stuttgart, Germany E-mail: [email protected] Background: Besides obesity and insulin resistance an increased translocation of intestinal bacterial endotoxin seems to be a risk factor for the development of nonalcoholic steatohepatitis (NASH). Aim: The aim of the present study was to investigate if an oral supplementation with Bifidobacterium adolescentis (B.a.) protects against diet-induced steatohepatitis. Method: C57BL/6 mice were either fed with a western style or control diet for 12 weeks ad libitum. Half of the mice received B.a. (107 cfu/ml) on a randomised basis. Liver injury and LPSconcentration in portal blood were determined after 12 weeks. Expression of MyD88, TNF-a and TLR-4 mRNA was measured using real time PCR. Furthermore, TLR-4 protein, neutrophil and 4-hydroxynonenale protein adducts were examined in paraffinembedded liver sections. Results: Mice fed a western style diet gained significant more weight than mice fed a control diet and developed a beginning steatohepatitis. Treatment with western style diet and B.a. supplementation significantly attenuated liver damage (inflammation counts dropped from 1.0 to 0.4, p < 0.05) compaired to western style groups without B.a. treatment. The protective effects of B.a. supplementation in the western style fed mice were associated with lower TLR-4 protein levels, number of infiltrating neutrophils and formation of 4-hydroxynonenal adducts as well as
Journal of Hepatology 2012 vol. 56 | S389–S548
S497
POSTERS lower MyD88, TLR-4 and TNF-a mRNA expression. However, portal endotoxin levels did not differ following B.a. treatment. Conclusion: These data suggest that 1. an oral B.a. supplementation can attenuate diet-induced steatohepatitis in a mouse model and 2. the protective effect is associated with a modulation of the TLR-4 signalling cascade in the liver. 1259 THE LIVER REDOX STATUS IN AN ANIMAL MODEL OF ESTROGEN DEFICIENCY ASSOCIATED WITH RENOVASCULAR HYPERTENSION AND THE BENEFICIAL EFFECTS OF TIBOLONE E.H. Gilglioni1 , L.B. Campos1 , C.R. Ambiel1 , M.N. Brito1 , R.F. Garcia1 , E.L. Ishii-Iwamoto2 , C.L. Salgueiro-Pagadigorria1 . 1 Physiological Sciences, 2 Biochemistry, University of Maringa, Maringa, Brazil E-mail: [email protected] Aims and Background: Hypertension is a frequent condition in post-menopausal women. This work was planned to evaluate the liver redox status in an animal model of oestrogen deficiency associated with renovascular hypertension (two-kidneys, one clip, 2K1C), and the possibility of beneficial effects of tibolone, a drug that has been suggested to be effective in treating many menopause symptoms. Methods: Ovariectomized hypertensive (OVX + 2K1C) Wistar rats were used in this study. The results were compared to those obtained with ovariectomized (OVX) and control (sham-operated) rats. Three weeks after the surgical procedures, rats of OVX + 2K1C group received daily doses of tibolone (0.04 mg/kg – OVX + 2K1CT rats) or vehicle (OVX + 2K1C), over a period of 15 days. Thereafter, the rats were anaesthetized for direct blood pressure measurements and liver removal. The liver redox status was evaluated through measurements of reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS) contents, as well as the activities of five antioxidants enzymes and the mitochondrial reactive oxygen species (ROS) generation were evaluated. Results: OVX + 2K1C rats presented mean blood pressure significantly higher (+28%), as compared to control and OVX rats. The treatment with tibolone (OVX + 2K1CT) reduced the blood pressures to values close to ones found in control and OVX rats. The livers content of GSH was increased and of TBARS was decreased in the same group of animals (OVX + 2K1CT) as compared with all other groups. The rates of mitochondrial ROS generation, which were increased in OVX + 2K1C rats (+52%), decreased in OVX + 2K1CT rats to values similar to those found in control and OVX rats. Among the antioxidant enzymes studied, stand out the beneficial effect of tibolone on the glucose-6-phosphate dehydrogenase activity, which was significantly reduced in OVX and OVX + 2K1C rats, and was reestablished in OVX + 2K1CT to values similar to control rats. Conclusion: Tibolone was effective in reducing the mean blood pressure in OVX + 2K1CT rats and this effect was accompanied of beneficial effects on liver redox status. Acknowledgements: This work was supported by Fundac˜ ¸ ao Araucaria ´ and grants from CNPq. 1260 AGING PROMOTES HEPATOCELLULAR STEATOSIS, INFLAMMATION AND FIBROSIS IN NON-ALCOHOLIC STEATOHEPATITIS IN MICE M. Saugspier, C. Dorn, C. Hellerbrand. University Hospital Regensburg, Regensburg, Germany E-mail: [email protected] Nonalcoholic steatohepatitis (NASH) involves necroinflammatory activity and fibrosis, which are believed to be mediated by lipotoxicity caused by toxic metabolites from excess fatty acids. Epidemiological studies indicate that age has impact on the natural course of NAFLD. S498
The aim of this study was to assess whether there are age dependent differences in the susceptibility to lipotoxicity and NASH in primary hepatocytes and a dietary murine model, respectively. Methods and Results: Upon stimulation with free fatty acids hepatocytes isolated from 4 week old mice revealed a significantly higher intracellular lipid accumulation and pro-inflammatory gene expression than hepatocytes isolated from 14 week old mice. Furthermore, we started feeding a high fat diet (HFD) to male C57BL/6 mice at the age of 4 or 14 weeks. After 12 weeks HFDfeeding we observed a significant increase of body weight and liver-to-body weight ratio as well hepatic triglyceride and freefatty-acid levels and histological steatosis in mice of both age classes as compared to control mice fed with standard chow. Furthermore, hydroxynonenal staining and increased Ncf-1 and Nox2 expression were indicative for oxidative stress. However, all these changes were significantly higher in the old mice. Similarly, serum transaminase levels, hepatic pro-inflammatory (TNF, IL-1, MCP-1) and pro-fibrogenic (TGF-beta, TIMP-1, Collagen I) gene expression, activation of hepatic stellate cells (evidenced by alphasma expression), histological matrix deposition and fibrosis were significantly higher in the older mice. Expression profiling applying Affymetrix technology revealed significant differences in critical enzymes involved in lipogenesis and lipid combustion between young and old mice. Conclusion: There is significant age-dependent variation in the susceptibility to NASH development and progression, which seems to be caused at least in part by differences in the lipid metabolisms in the hepatocytes. The identification of the underlying (molecular) mechanisms of these differences may lead to new prognostic markers or novel therapeutic targets for the progression of NAFLD. 1261 MICE FED A HIGH-FAT HIGH-FRUCTOSE DIET AND TREATED WITH LOW DOSE STREPTOZOTOCIN DEVELOP INSULIN RESISTANCE, NASH AND LIVER FIBROSIS Y.O. Kim, M. Stoll, B. Hebich, X. Wang, D. Schuppan. Molecular and Translational Medicine, Medicine I./Univ. Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany E-mail: [email protected] Background and Aims: Despite recent improvements, an optimal model that reflects all features of human non-alcoholic steatohepatitis (NASH) is needed. We developed a NASH model that combines a high-fructose/high fat diet with low dose Streptozotocin (STZ) to induce hyperglycemia with hyperinsulinemia but relative insulin resistance. Methods: Six week old female C57BL/6 mice were randomly assigned to standard chow (13% kcal from fat; 65% kcal from complex carbohydrates, n = 10), or to a high-fat and highcarbohydrate diet (HF/HC; Surwit: 59% kcal fat; 26% kcal complex carbohydrates, plus 55% fructose and 45% sucrose in drinking water; n = 20) for 6, 12 and 18 weeks. 10/20 mice on the HF/HC diet received low dose STZ i.p. (HF/HC/STZ group; 65 mg STZ/kg; four weeks before sacrifice on two consecutive days) to induce relative insulin deficiency despite hyperinsulinemia, as found in type 2 diabetes. Parameters of insulin resistance and liver function, intrahepatic lipid, inflammation, fibrosis (hydroxyproline, Sirius red morphometry) and transcript levels related to fibrogenesis and fibrolysis (qPCR) were determined at sacrifice. Results: Compared to animals on normal chow, mice on HF/HC and HF/HC/STZ showed significantly more weight gain (up to 15 g at 18 w) and significantly higher serum markers of insulin resistance compared with normal chow (HOMA score, elevated 1.6-fold at 12 w). While the HF/HC and the HF/HC/STZ groups developed comparable increases of liver weight (1.5-fold) and fat (29%) compared with normal chow, collagen content of the HF/HC/STZ group was significantly higher than in the HF/HC group at week
Journal of Hepatology 2012 vol. 56 | S389–S548
intracellular vesicles to the cell surface, thus mediating insulindependent glucose uptake. Based on its several metabolic roles, we hypothesized that sortilin affects whole body glucose metabolism and hepatic steatosis and inflammation. With respect to hepatic fat accumulation, it has been shown that increased lipogenesis which ends up with triglyceride accumulation serves as a favorable fat store protecting the liver from inflammation. Methods: C57BL/6 (WT) and sortilin −/− male mice were fed regular chow (RC) and Western diet for two months and methioninecholine deficient diet (MCD) for 5 weeks. Results: Sortilin−/− mice have significantly reduced body weight and visceral fat both after RC and Western diet, despite similar food intake. In addition, sortilin−/− mice have significantly lower serum cholesterol and exhibit better glucose tolerance tests. Interestingly, hepatic lipogenesis was increased, as demonstrated by increased mRNA expression of fatty acid synthase and stearoyl CoA desaturase. Hepatic expression of phosphorylated acetyl CoA carboxylase was significantly lower in sortilin−/− mice, also indicative of increased lipogenesis. In addition, livers of sortilin −/− mice showed reduced mRNA levels of hepatic inflammatory, insulin resistance-inducing cytokines TNFa and PAI-1. Adipose tissue of sortilin−/− displays significantly smaller adipocyte size and decreased mRNA levels of inflammatory cytokines TNFa, PAI1and MCP-1. Muscle tissue of these mice express higher levels of glut4 and differentiation marker myogenin. These data suggest that sortilin −/− have better metabolic parameters and attenuated liver inflammation, even though they exhibit increased hepatic lipogenesis. In accordance with the above, sortilin−/− mice fed MCD show attenuated liver damage as expressed by 5-fold lower AST (143 vs 875unit/L) and twice lower ALT (132 vs 282unit/L), yet stronger steatosis. Conclusion: Lack of sortilin induces a beneficial metabolic phenotype and attenuated liver inflammation in metabolic stress models, accompanied by increased lipogenesis. 1258 ORAL BIFIDOBACTERIUM ADOLESCENTIS SUPPLEMENTATION ATTENUATES NONALCOHOLIC STEATOHEPATITIS (NASH) IN A MOUSE MODEL A. Reichold, S. Brenner, A. Spruss, A. Rings, K. Forster-Fromme, ¨ I. Bergheim, S.C. Bischoff. Department of Nutritional Medicine (180a), University of Hohenheim, Stuttgart, Germany E-mail: [email protected] Background: Besides obesity and insulin resistance an increased translocation of intestinal bacterial endotoxin seems to be a risk factor for the development of nonalcoholic steatohepatitis (NASH). Aim: The aim of the present study was to investigate if an oral supplementation with Bifidobacterium adolescentis (B.a.) protects against diet-induced steatohepatitis. Method: C57BL/6 mice were either fed with a western style or control diet for 12 weeks ad libitum. Half of the mice received B.a. (107 cfu/ml) on a randomised basis. Liver injury and LPSconcentration in portal blood were determined after 12 weeks. Expression of MyD88, TNF-a and TLR-4 mRNA was measured using real time PCR. Furthermore, TLR-4 protein, neutrophil and 4-hydroxynonenale protein adducts were examined in paraffinembedded liver sections. Results: Mice fed a western style diet gained significant more weight than mice fed a control diet and developed a beginning steatohepatitis. Treatment with western style diet and B.a. supplementation significantly attenuated liver damage (inflammation counts dropped from 1.0 to 0.4, p < 0.05) compaired to western style groups without B.a. treatment. The protective effects of B.a. supplementation in the western style fed mice were associated with lower TLR-4 protein levels, number of infiltrating neutrophils and formation of 4-hydroxynonenal adducts as well as
Journal of Hepatology 2012 vol. 56 | S389–S548
S497
POSTERS lower MyD88, TLR-4 and TNF-a mRNA expression. However, portal endotoxin levels did not differ following B.a. treatment. Conclusion: These data suggest that 1. an oral B.a. supplementation can attenuate diet-induced steatohepatitis in a mouse model and 2. the protective effect is associated with a modulation of the TLR-4 signalling cascade in the liver. 1259 THE LIVER REDOX STATUS IN AN ANIMAL MODEL OF ESTROGEN DEFICIENCY ASSOCIATED WITH RENOVASCULAR HYPERTENSION AND THE BENEFICIAL EFFECTS OF TIBOLONE E.H. Gilglioni1 , L.B. Campos1 , C.R. Ambiel1 , M.N. Brito1 , R.F. Garcia1 , E.L. Ishii-Iwamoto2 , C.L. Salgueiro-Pagadigorria1 . 1 Physiological Sciences, 2 Biochemistry, University of Maringa, Maringa, Brazil E-mail: [email protected] Aims and Background: Hypertension is a frequent condition in post-menopausal women. This work was planned to evaluate the liver redox status in an animal model of oestrogen deficiency associated with renovascular hypertension (two-kidneys, one clip, 2K1C), and the possibility of beneficial effects of tibolone, a drug that has been suggested to be effective in treating many menopause symptoms. Methods: Ovariectomized hypertensive (OVX + 2K1C) Wistar rats were used in this study. The results were compared to those obtained with ovariectomized (OVX) and control (sham-operated) rats. Three weeks after the surgical procedures, rats of OVX + 2K1C group received daily doses of tibolone (0.04 mg/kg – OVX + 2K1CT rats) or vehicle (OVX + 2K1C), over a period of 15 days. Thereafter, the rats were anaesthetized for direct blood pressure measurements and liver removal. The liver redox status was evaluated through measurements of reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS) contents, as well as the activities of five antioxidants enzymes and the mitochondrial reactive oxygen species (ROS) generation were evaluated. Results: OVX + 2K1C rats presented mean blood pressure significantly higher (+28%), as compared to control and OVX rats. The treatment with tibolone (OVX + 2K1CT) reduced the blood pressures to values close to ones found in control and OVX rats. The livers content of GSH was increased and of TBARS was decreased in the same group of animals (OVX + 2K1CT) as compared with all other groups. The rates of mitochondrial ROS generation, which were increased in OVX + 2K1C rats (+52%), decreased in OVX + 2K1CT rats to values similar to those found in control and OVX rats. Among the antioxidant enzymes studied, stand out the beneficial effect of tibolone on the glucose-6-phosphate dehydrogenase activity, which was significantly reduced in OVX and OVX + 2K1C rats, and was reestablished in OVX + 2K1CT to values similar to control rats. Conclusion: Tibolone was effective in reducing the mean blood pressure in OVX + 2K1CT rats and this effect was accompanied of beneficial effects on liver redox status. Acknowledgements: This work was supported by Fundac˜ ¸ ao Araucaria ´ and grants from CNPq. 1260 AGING PROMOTES HEPATOCELLULAR STEATOSIS, INFLAMMATION AND FIBROSIS IN NON-ALCOHOLIC STEATOHEPATITIS IN MICE M. Saugspier, C. Dorn, C. Hellerbrand. University Hospital Regensburg, Regensburg, Germany E-mail: [email protected] Nonalcoholic steatohepatitis (NASH) involves necroinflammatory activity and fibrosis, which are believed to be mediated by lipotoxicity caused by toxic metabolites from excess fatty acids. Epidemiological studies indicate that age has impact on the natural course of NAFLD. S498
The aim of this study was to assess whether there are age dependent differences in the susceptibility to lipotoxicity and NASH in primary hepatocytes and a dietary murine model, respectively. Methods and Results: Upon stimulation with free fatty acids hepatocytes isolated from 4 week old mice revealed a significantly higher intracellular lipid accumulation and pro-inflammatory gene expression than hepatocytes isolated from 14 week old mice. Furthermore, we started feeding a high fat diet (HFD) to male C57BL/6 mice at the age of 4 or 14 weeks. After 12 weeks HFDfeeding we observed a significant increase of body weight and liver-to-body weight ratio as well hepatic triglyceride and freefatty-acid levels and histological steatosis in mice of both age classes as compared to control mice fed with standard chow. Furthermore, hydroxynonenal staining and increased Ncf-1 and Nox2 expression were indicative for oxidative stress. However, all these changes were significantly higher in the old mice. Similarly, serum transaminase levels, hepatic pro-inflammatory (TNF, IL-1, MCP-1) and pro-fibrogenic (TGF-beta, TIMP-1, Collagen I) gene expression, activation of hepatic stellate cells (evidenced by alphasma expression), histological matrix deposition and fibrosis were significantly higher in the older mice. Expression profiling applying Affymetrix technology revealed significant differences in critical enzymes involved in lipogenesis and lipid combustion between young and old mice. Conclusion: There is significant age-dependent variation in the susceptibility to NASH development and progression, which seems to be caused at least in part by differences in the lipid metabolisms in the hepatocytes. The identification of the underlying (molecular) mechanisms of these differences may lead to new prognostic markers or novel therapeutic targets for the progression of NAFLD. 1261 MICE FED A HIGH-FAT HIGH-FRUCTOSE DIET AND TREATED WITH LOW DOSE STREPTOZOTOCIN DEVELOP INSULIN RESISTANCE, NASH AND LIVER FIBROSIS Y.O. Kim, M. Stoll, B. Hebich, X. Wang, D. Schuppan. Molecular and Translational Medicine, Medicine I./Univ. Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany E-mail: [email protected] Background and Aims: Despite recent improvements, an optimal model that reflects all features of human non-alcoholic steatohepatitis (NASH) is needed. We developed a NASH model that combines a high-fructose/high fat diet with low dose Streptozotocin (STZ) to induce hyperglycemia with hyperinsulinemia but relative insulin resistance. Methods: Six week old female C57BL/6 mice were randomly assigned to standard chow (13% kcal from fat; 65% kcal from complex carbohydrates, n = 10), or to a high-fat and highcarbohydrate diet (HF/HC; Surwit: 59% kcal fat; 26% kcal complex carbohydrates, plus 55% fructose and 45% sucrose in drinking water; n = 20) for 6, 12 and 18 weeks. 10/20 mice on the HF/HC diet received low dose STZ i.p. (HF/HC/STZ group; 65 mg STZ/kg; four weeks before sacrifice on two consecutive days) to induce relative insulin deficiency despite hyperinsulinemia, as found in type 2 diabetes. Parameters of insulin resistance and liver function, intrahepatic lipid, inflammation, fibrosis (hydroxyproline, Sirius red morphometry) and transcript levels related to fibrogenesis and fibrolysis (qPCR) were determined at sacrifice. Results: Compared to animals on normal chow, mice on HF/HC and HF/HC/STZ showed significantly more weight gain (up to 15 g at 18 w) and significantly higher serum markers of insulin resistance compared with normal chow (HOMA score, elevated 1.6-fold at 12 w). While the HF/HC and the HF/HC/STZ groups developed comparable increases of liver weight (1.5-fold) and fat (29%) compared with normal chow, collagen content of the HF/HC/STZ group was significantly higher than in the HF/HC group at week
Journal of Hepatology 2012 vol. 56 | S389–S548