泽泻汤通过ROS/mTOR通路抑制自噬干预NASH肝纤维化的机制研究

氧化应激诱导的自噬是NASH肝纤维化主要发病机制之一。本项目组前期临床研究发现，具有健脾泄浊，化湿利水功效的泽泻汤可以改善非酒精性脂肪肝患者的肝纤维化指标，进一步实验发现，泽泻汤改善NASH肝纤维化模型小鼠的肝纤维化病变，同时，在电镜下可见泽泻汤干预组自噬小体数明显减少，小鼠肝脏α-SMA及LC3-Ⅱ表达下调，p-P70S6K表达上调，而泽泻汤可以显著降低机体的氧化应激性，据此我们推测泽泻汤可能通过ROS/mTOR通路抑制自噬从而发挥抗肝纤维作用。本课题组拟构建体内外NASH肝纤维化模型，观察泽泻汤的抗氧化作用，通过电镜及MDC染色法结合荧光显微技术观察自噬通路的变化，并观察ROS/mTOR及自噬通路信号分子的表达变化，运用分子生物学方法阐释泽泻汤干预NASH肝纤维化的分子机制，为中西医结合治疗NASH肝纤维化发现新的候选靶点和实验依据。

Oxidative stress induced autophagy is one of the main pathogenesis of NASH hepatic fibrosis. Preliminary clinical study found that Zexie decoction can improve nonalcoholic fatty liver in patients with liver fibrosis, further study found that Zexie Decoction improve hepatic fibrosis, hepatic fibrosis in NASH mice, at the same time, in the electron microscope of Zexie Decoction intervention group significantly reduced the number of autophagosomes, down regulate the expression of mouse liver alpha -SMA and LC3- II. Expression of p-P70S6K, and Zexie decoction can significantly reduce the oxidative stress of the body, so we speculate that Zexie decoction may be through the ROS/mTOR pathway and inhibition of autophagy can play with anti hepatic fibrosis. The research group will build a classic NASH in vitro and in vivo hepatic fibrosis model, to observe the antioxidation effect of Zexie decoction, combined with the changes of autophagy pathway were observed by electron microscopy and fluorescence microscopy with MDC staining method, and observe the expression of ROS/mTOR and autophagy signaling pathway, using molecular biology method to explain the molecular mechanism of Zexie Decoction intervention of NASH hepatic fibrosis combined treatment of liver fibrosis, to find a new candidate target and experimental basis for the treatment of NASH hepatic fibrosis with integrated traditional Chinese and Western Medicine.