大黄酸联合吡格列酮对大鼠非酒精性脂肪性肝炎治疗作用机制的探讨

Effects of Combination Therapy of Rehin and Pioglitazone on Rats with Nonalcoholic Steatohepatitis

关键词:  非酒精性脂肪性肝病  大黄酸  吡格列酮  胰岛素抵抗  脂质代谢紊乱

Key words:  nonalcoholic steatohepatitis  Rhein  Pioglitazone  insulin resistance  lipid metabolism disorder

摘要:

目的 探讨大黄酸联合吡格列酮对高脂诱建的大鼠NASH协同治疗及其可能的作用机制。方法 清洁级SD大鼠 140~160 g，♂，76 只正常喂养1 周后，随机分成6 组(对照组、模型组各为14只，余组均为12只)：对照组，模型组，大黄酸组，吡格列酮组，大黄酸吡格列酮联合低剂量组，高剂量组。模型组高脂高胆固醇饲料由普通饲料+10%猪油+2%胆固醇配制，药物组均在12周高脂饮食后即造模成功后给予干预。大黄酸用生理盐水配成5 g·L^-1混悬液，每天固定时间100 mg·kg^-1·d^-1灌胃，吡格列酮组8 mg·kg^-1·d^-1，联合低剂量组取其相对有效低剂量，大黄酸50 mg·kg^-1·d^-1，吡格列酮组4 mg·kg^-1·d^-1。联合高剂量组为大黄酸100 mg·kg^-1·d^-1，吡格列酮组8 mg·kg^-1·d^-1。于第20 周处死。所有动物测体重、肝湿重，计算肝指数；测空腹血糖、转氨酶及血脂水平，放免法测空腹胰岛素及TNF-a，计算胰岛素抵抗指数；酶法测定肝组织匀浆MDA、GSH-PX水平；HE 染色肝病理组织切片。结果 第20 周模型组大鼠血糖、胰岛素、血脂、ALT、AST及胰岛素抵抗指数较对照组明显增高；肝组织出现重度脂肪变性，均出现小叶内炎症细胞浸润和散在的灶性坏死；药物组大鼠的生化指标和脂肪变及炎症程度较模型组均有明显减轻，联合组较药物单用组更为有效。结论 大黄酸联合吡格列酮较两药单用能更有效抑制脂质过氧化反应及改善胰岛素抵抗，对大鼠NASH 具有明显的协同治疗作用。

Abstract:

OBJECTIVE To elucidate effects of combination therapy of rehin and Pioglitazone on nonalcoholic steatohepatitis in rats with NASH and investigate the synergic mechanism of rehin and Pioglitazone on NASH. METHODS Totally 76 male SD rats were randomly divided into 6 groups after fed with normal diet for 7 days: Control group (n=14): fed with normal diet for 20 weeks (2 rats for 12 weeks and then executed); Model group (n=14): fed with high-fat and high-cholesterol diet consisting of the normal diet, 10% lard and 2% cholesterol, for 20 weeks (2 rats for 12 weeks and then were executed);Rhein group (n=12): Rhein was added orally after 12 weeks fed with high-fat and high-cholesterol diet;Pioglitazone group (n=12): Pioglitazone was added orally after 12 weeks fed with high-fat and high-cholesterol diet;combination group were divided into low-dose and high-dose group(n=12): different doses of Rhein and Pioglitazone were added orally after 12 weeks fed with high-fat and high-cholesterol diet. All the rats were executed after 20 weeks. Body mass, liver weight, transaminase level, blood glucose, triglyceride, cholesteral, insulin level and liver histology were detected. Serum insulin and tumor necrosis factor-a(TNF-a) level were measured by radio-immunity technique. The level of malondialdehyde (MDA), glutathione peroxidase (GSH-PX) was measured by chromatometry. RESULTS After 20 weeks, the model group developed lipid metabolic disorder and insulin resistance. Livers presented the pathology of severe hepatocyte steatosis and the sporadic inflammatory cell infiltration or focal necrosis. Lipid metabolic disorder and insulin resistance in intervention group were improved, along with the status of hepatocyte steatosis and inflammatory cell infiltration, compared with model group. Combination group was more effective than the single drug group. CONCLUSION Combination therapy of rehin and Pioglitazone can prevent the development of nonalcoholic steatohepatitis and is more effective than the single drug group. Combination group has obvious synergic therapeutic effect.