基于miR-181c-5p介导肝细胞线粒体损伤研究疏肝消脂方干预NASH伴肝纤维化的作用机制

线粒体损伤在非酒精性脂肪性肝炎（NASH）伴纤维化发展过程中起关键的作用，但是肝脂肪变性所损伤的线粒体导致肝组织炎症伴纤维化的过程，是否受microRNA调节仍不清楚。前期研究发现疏肝消脂方（SX）能抗肝炎与纤维化、并对miR-181c-5p异常表达有明显调控作用，据此提出“SX通过作用于核基因编码的miR-181c-5p，调控肝细胞线粒体呼吸链(MRC） Cox1、Cox2和Cox3 mRNA和蛋白表达，减轻其功能障碍，干预脂肪变性、炎症、纤维形成过程，产生对NASH伴肝纤维化的治疗作用”假说。用qRT-PCR、NB、LNA-FISH、CRISPR/Cas9等技术，在体内外观察：①SX对NASH伴肝纤维化MRC的影响;②SX对NASH伴肝纤维化线粒体miR-181c-5p靶向调节机制。明确miR-181c-5p对肝细胞MRC复合物功能调控作用，以及SX治疗NASH伴肝纤维化的作用靶点。

Mitochondrial dysfunction plays a key role in the development of non-alcoholic steatohepatitis (NASH) with fibrosis, but whether microRNA regulates the process of liver inflammation with fibrosis caused by mitochondria dysfunction by hepatic steatosis remains unclear. Our previous studies have found that Shugan Xiaozhi Decoction (SX) can anti-hepatic inflammation, anti-liver fibrosis. Moreover, SX has a significant regulatory effect on the abnormal expression of miR-181c-5p. We hypothesize that "SX regulates the mRNA and protein expressions of the Cox1, Cox2 and Cox3 of hepatocytic mitochondrial respiratory chain (MRC) by acting on the intracellular translocation of miR-181c-5p which is encoded by nuclear DNA, alleviate the mitochondrial dysfunction of hepatocytes, intervene in liver steatosis, inflammation and fibrogenesis, and produce therapeutic effects on NASH with fibrosis". qRT-PCR、NB、LNA-FISH、CRISPR/Cas9 and other methods will be used in vivo and in vitro to observe ① Effects of SX on MRC in NASH with fibrosis ② The mechanism of targeted modulation of mitochondrial miR-181c-5p in NASH with fibrosis by SX. Our objective is to clarify the role of miR-181c-5p in the regulation of the function of MRC complex in hepatocytes, and to elucidate the therapeutic target of SX for NASH with fibrosis.