抗脂方改善大鼠非酒精性脂肪性肝炎的实验研究

Amelioration effects of Kangzhi Recipe on nonalcoholic steatohepatitis in rats

目的研究中药抗脂方调节糖脂代谢,改善非酒精性脂肪性肝炎的作用。方法将SD大鼠随机分为正常对照组(A组)、脂肪性肝炎模型组(B组)、脂肪性肝炎给予易善复组(C组)和脂肪性肝炎给予抗脂方组(D组)。除正常对照组外,其余各组以高脂饲料喂养,同时皮下注射CCl4造模。造模同时灌胃给药8周。以常规方法检测肝功能、生化指标、肝组织病理染色和HOMA-IR;采用ELISA法检测脂联素(ADP/Acrp3)、过氧化物酶体增殖物激活受体α(PPAR-α)、肿瘤坏死因子-α(TNF-α)、空腹胰岛素(FINS)、游离脂肪酸(FFA)和蛋白酪氨酸激酶(PTK);用Western blotting法检测胰岛素信号通路相关蛋白表达。结果与脂肪性肝炎模型组比较,脂肪性肝炎给予抗脂方组明显改善了非酒精性脂肪性肝炎大鼠的肝功能,逆转了其相关的病理特征及生化指数,差异有统计学意义(P<0.05,P<0.01);同时,脂肪性肝炎给予抗脂方组大鼠FPG、FINS、HOMA-IR降低(P<0.01),PTK、PPAR-α、ADP升高(P<0.01),TNF-α降低(P<0.01),胰岛素受体IR增加(P<0.01),IRS1、IRS2升高(P<0.05),PI3K表达及Akt的磷酸化表达明显升高(P<0.05),p38的磷酸化水平明显降低(P<0.01),从而增加了大鼠肝细胞对胰岛素的敏感性,改善了胰岛素信号转导。结论中药抗脂方具有明显改善非酒精性脂肪性肝炎的作用,其相关的机制可能与抗脂方调节肝脏的糖脂代谢、改善胰岛素信号通路有关。

Objective To study the effects of Kangzhi Recipe on regulating glucose and lipid metabolism and improving nonalcoholic steatohepatitis. Methods SD rats were randomly divided into the normal control group,model group of steatohepatitis,Yishanfu Capsule（ polyene phosphatidylcholine capsules） group and Kangzhi Recipe group.Except the normal control group,the other groups were fed with high fat diet and treated with CCl4 by subcutaneous injection to establish the model. During the modeling,each group was treated with the corresponding drugs by intragastric administration for 8 weeks. The liver function,biochemical indexes,pathological staining of liver tissue and homeostasis model assessment for insulin resistance（ HOMA-IR） were routinely detected. The levels of adiponectin（ ADP/Acrp3）,peroxisome proliferator-activated receptor-α（ PPAR-α）,tumor necrosis factor-α（ TNF-α）,fasting insulin（ FINS）,free fatty acid（ FFA） and protein tyrosine kinase（ PTK） were detected by ELISA. The expressions of related proteins in insulin signaling pathway were detected by Western blotting. Results Compared with the model group of steatohepatitis,the liver function of rats with nonalcoholic steatohepatitis were obviously improved in the Kangzhi Recipe group,and the related pathological characteristics and biochemical indexes were reversed,with statistically significant differences（ P〈0. 05,P〈0. 01）. And after the treatment of Kangzhi Recipe,the levels of fasting plasma glucose（ FPG）,FINS and HOMA-IR were decreased（ P〈0. 01）,the contents of PTK,PPAR-α and ADP were increased（ P〈0. 01） and the content of TNF-α was decreased（ P〈0. 01）,the protein expression of insulin receptor（ IR） was increased（ P〈0. 01） and the protein expressions of insulin receptor substrate 1（ IRS1） and IRS2 were increased（ P〈0. 05）,the expressions of PI3K and phosphorylated Akt were obviously increased（ P〈0. 05） and the expression of phosphorylated p38 was obviously decreased（ P〈0. 01）. It suggested that the sensitivity of rat hepatocytes to insulin was increased and the insulin signaling transduction was improved. Conclusion Kangzhi Recipe can significantly improve nonalcoholic steatohepatitis. Its mechanism may be associated with the regulation on hepatic glucose and lipid metabolism and improvement on insulin signaling pathway.