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目的探讨温运清利法对非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)大鼠肝组织甘油二酯(diacylglycerol,DAG)/蛋白激酶Cε(protein kinase Cε,PKCε)信号通路的影响。方法高脂饮食饲养SD大鼠8周制备NASH模型,造模成功后,将大鼠随机分为模型组、苓桂术甘汤组、茵陈蒿汤组、合方组(苓桂术等与茵陈蒿汤)、罗格列酮组,共5组,每组10只;另有空白组10只。药物治疗4周后,运用自动生化分析仪检测血清生化指标,HE染色、油红O染色观察肝组织病理,ELISA法检测肝组织DAG,RT-PCR检测PKCεmRNA、TNF-αmRNA,Western blot检测PKCε和TNF-α蛋白的表达。结果 (1)各给药组大鼠一般状况、肝组织脂肪变性和炎症反应程度、血清生化指标等均较模型组有一定的减轻;(2)苓桂术甘汤、合方组、罗格列酮组大鼠肝组织DAG较模型组明显降低(P<0.05,P<0.01);(3)各治疗组大鼠肝组织PKCεmRNA和蛋白含量均显著降低(P<0.05,P<0.01),茵陈蒿汤组、合方组及罗格列酮组大鼠肝组织TNF-αmRNA和蛋白含量显著降低(P<0.05)。结论温运清利法可能是通过下调肝组织DAG,抑制PKCε表达,降低TNF-α水平,从而对肝内脂质代谢、胰岛素抵抗、炎症损伤起到了调节作用,DAG/PKCε通路有可能成为治疗NASH的新靶点。
Objective To investigate the effects of warming, clearing and thawing on hepatic diacylglycerol (DAG) / protein kinase C (PKCε) signal pathways in non-alcoholic steatohepatitis (NASH) rats. Methods SD rats were fed with high-fat diet for 8 weeks to prepare NASH model. After successful modeling, the rats were randomly divided into model group, Lingguizhugan decoction group, Yinchenhao Decoction group and Hefang decoction group Artemisia soup), rosiglitazone group, a total of 5 groups of 10; another blank group of 10. Four weeks after the drug treatment, serum biochemical indexes were detected by automatic biochemical analyzer. Liver pathology was observed by HE staining and oil red O staining. DAG in liver tissue was detected by ELISA. PKCε mRNA and TNF-α mRNA were detected by RT-PCR, TNF-α protein expression. Results (1) The general condition, the degree of hepatic steatosis, the degree of inflammatory reaction and the serum biochemical indexes of the rats in each administration group were somewhat relieved compared with the model group. (2) The rats in Lingguizhugan decoction, Hefang group, Compared with model group, the DAG in liver of rats in ketoprofen group was significantly lower than that in model group (P <0.05, P <0.01); (3) The content of PKCεmRNA and protein in liver tissue of rats in each group were significantly decreased The levels of TNF-αmRNA and protein in the liver of the Chenhao decoction group, the combination group and the rosiglitazone group were significantly decreased (P <0.05). Conclusion Wen-Wen Qing-Li method may regulate the intrahepatic lipid metabolism, insulin resistance and inflammation damage by down-regulating DAG, inhibiting the expression of PKCε and decreasing the level of TNF-α, and the DAG / PKCε pathway may be the treatment of NASH New target