加味茵陈五苓散对非酒精性脂肪肝模型大鼠miR-34a及糖异生的影响

Effects of modified Yinchen Wuling Powder on miR-34a and gluconeogenesis in non-alcoholic fatty liver model rats

关键词:  加味茵陈五苓散  非酒精性脂肪肝  miR-34a  过氧化物酶体增殖物激活受体γ共激活因子-α  磷酸烯醇式丙酮酸羧激酶

Key words:  modified YinchenWuling Powder  NAFLD  miR-34a  PGC-1α  PEPCK

摘要:

[目的] 探讨加味茵陈五苓散对非酒精性脂肪肝（NAFLD）模型大鼠miR-34a、过氧化物酶体增殖物激活受体-γ共激活因子-1α（PGC-1α）、磷酸烯醇式丙酮酸羧激酶（PEPCK）表达的影响。[方法] 将50只SD大鼠随机分为5组：正常组、模型组、中药组、中药+空载体病毒组、中药+慢病毒组。高脂饲料喂养10周复制NAFLD大鼠模型，并应用慢病毒过表达miR-34a，在光镜下苏木精-伊红（HE）染色观察肝细胞结构及脂肪含量；全自动生化分析仪检测血清天门冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）、总胆固醇（TC）、三酰甘油（TG）水平；定时定量荧光聚合酶链反应（Realtime PCR）检测大鼠肝脏miR-34a、PGC-1α、PEPCK mRNA的表达。[结果] 与正常组比较，模型组出现明显肝脏脂肪变性；血清ALT、AST、TC、TG水平明显升高（P<0.05）；相较于模型组，中药组及中药+空载体病毒组肝内脂肪变性明显减轻（P<0.05），血清ALT、AST、TC、TG水平均降低（P<0.05）。miR-34a、PGC-1α、PEPCK表达情况：与正常组比较，模型组miR-34a表达明显增高（P<0.05），PGC-1α和PEPCK表达降低；使用中药干预后，与模型组比较，中药组及中药+空载体病毒组miR-34a表达明显降低（P<0.05），PGC-1α和PEPCK表达明显增高（P<0.05），中药+慢病毒组miR-34a、PGC-1α和PEPCK的表达无明显变化（P>0.05）。[结论] 加味茵陈五苓散能够改善改善肝脂肪变性，可能与降低miR-34a的表达、进而提高PGC-1α和PEPCK表达，促进糖异生。

Abstract:

[Objective] To observe the effects of modified Yinchen Wuling Powder on the expression of miR-34a,PGC-1α and PEPCK in Nonalcoholic fatty liver (NAFLD) model rats.[Methods] Fifty SD rats were randomly divided into normal group,model group,Chinese medicine group,Chinese medicine+empty virus group and Chinese medicine+lentivirus group. Non-alcoholic fatty liver model rats were induced by a high-quality diet and miR-34a was overexpressed by lentivirus. The histological structure of HE staining liver tissue was observed under light microscope. The serum levels of AST,ALT,TC and TG were detected by automatic biochemical analyzer. The expression of miR-34a,PGC-1αand PEPCK mRNA weredetectedbyRealtime PCR.[Results] Compared with the normal group,the model group showed obvious hepatic steatosis. Serum levels of ALT,AST,TC and TG were also higher than those in normal group (P<0.05). Compared with the model group,intrahepatic steatosis was significantly reduced in Chinese medicine group and Chinese medicine+empty virus group (P<0.05),and serum levels of ALT,AST,TC and TG were all reduced (P<0.05). Expressions of miR-34a,PGC-1αand PEPCK:Compared with the normal group,the expression of miR-34a in model group was significantly increased (P<0.05),and the expression of PGC-1α in the model group was decreased(P<0.05). After intervention with Chinese medicine,compared with the model group,the expressions of miR-34a in Chinese medicine group and the Chinese medicine+empty virus group weresignificantly reduced (P<0.05),and the expression of PGC-1αand PEPCK were significantly increased (P<0.05),while the expression of miR-34a,PGC-1α and PEPCK in Chinese medicine+lentivirus group was not significantly changed (P>0.05).[Conclusion] Modified Yinchen Wuling Powder might promote mitochondrial biosynthesis to improve hepatic steatosisby reducing the expression of miR-34a and increasing the expression of PGC-1α and PEPCK.