降脂颗粒上调Atg5表达活化自噬治疗非酒精性脂肪性肝炎的机制研究

非酒精性脂肪性肝炎(NASH)发病广泛，机制不明，危害严重。研究表明，自噬活化改善脂代谢紊乱，也可调节脂肪变性情况下的肝细胞损伤。我们前期研究发现脂肪肝有效治疗药物降脂颗粒可诱导自噬活化并可上调自噬相关基因Atg5的表达。因此，我们提出“降脂颗粒通过上调Atg5表达活化自噬，抑制脂肪变和炎症治疗NASH”的科学假说。为验证该假说，本项目将采用饮食诱导的NASH小鼠模型，评价降脂颗粒对NASH的改善作用以及对自噬应答的影响；体外实验通过荧光标记、透射电镜、自噬阻诱导剂和阻断剂的应用评价药物对自噬状态和Atg5的调控；应用CRISPR-Cas9、慢病毒转染研究药物效应是否依赖于Atg5的表达，从而阐明降脂颗粒改善脂代谢和炎症治疗NASH的作用机理，进一步促进NASH新药研发和临床应用。

Non-alcoholic steatohepatitis (NASH) is with a wide range of incidence, unknown mechanisms and can lead both hepatic and metabolic risks to human being. Studies have shown that autophagy activation could improve dyslipidemia and hepatocellular steatosis and injury. Our previous studies found that Jiangzhi Granula, a formula targeting fatty liver diseases can induce autophagy and up-regulate the expression of autophagy-related gene Atg5. Therefore, we propose a scientific hypothesis: "Jiangzhi granule activates autophagy by up-regulating Atg5 expression, inhibits steatosis and inflammation in treating of NASH". To verify this hypothesis, diet-induced NASH mouse model will be used to evaluate the effect of Jiangzhi granule on NASH and its effect on autophagy. In vitro experiments would be carried out by applying fluorescence labeling, transmission electron microscopy, autophagy agonist and antagonist to detect the regulation of drug on autophagy and Atg5; Using CRISPR-Cas9, lentiviral transfection to study whether the drug effect depends on Atg5 expression, thus to clarify the mechanisms of Jiangzhi Granula on improving lipid metabolism and inflammation, and to further promote the NASH drug development and clinical applications.