参葛方通过AMPK-Insig1-SREBPs通路抑制肝细胞脂质沉积治疗非酒精性脂肪性肝炎机制研究

肝细胞脂质沉积是非酒精性脂肪性肝炎（NASH）发生发展的关键病理环节，AMPK磷酸化Insig1，增强其蛋白稳定性，从而抑制SREBPs活化入核（AMPK-Insig1-SREBPs通路），是抑制脂质合成的关键通路。前期研究证实参葛方能有效治疗NASH，减轻肝脏脂肪变。定量蛋白质组学发现，参葛方显著增加NASH小鼠肝脏Insig1表达，生物信息学分析提示，参葛方抑制肝细胞脂质沉积与AMPK-Insig1-SREBPs通路相关。由此提出假说：参葛方通过激活AMPK，促进Insig1磷酸化，抑制SREBPs活化，减少肝细胞脂质合成，治疗NASH。拟开展：①动物和细胞水平验证参葛方抑制肝细胞脂质沉积疗效。②结合体内、体外模型，采用AMPK抑制剂、RNA干扰和Insig1敲除小鼠，明确参葛方通过AMPK-Insig1-SREBPs通路抑制肝细胞脂质沉积。为进一步阐释参葛方治疗NASH提供理论依据。

Lipid accumulation in hepatocyte promotes the occurrence and development of non-alcoholic steatohepatitis (NASH). AMPK phosphorylates Insig1 which further inhibits the translocation of activated SREBPs into the nucleus (AMPK-Insig1-SREBPs pathway) is a key pathway to inhibit lipid synthesis. Previous studies have confirmed that Shenge Formula is effective for NASH and can reduce lipid accumulation in hepatocyte. Quantitative proteomics study revealed that Shenge Formula significantly increased the expression of Insig1 in the liver of mouse model. Bioinformatics analysis suggested that the inhibition of lipid accumulation by Shenge Formula was related to AMPK-Insig1-SREBPs pathway. Therefore, we proposed our hypothesis that “Shenge Formula promotes the phosphorylation of Insig1 by activating AMPK, which inhibits the activation of SREBPs, results in the inhibition of lipid synthesis in hepatocytes”. This project intends to combine in vivo and in vitro models, by using AMPK inhibitor, Insig1 RNA interference and Insig1 knockout mice to verify Shenge Formula inhibits lipid accumulation through activating AMPK-Insig1-SREBPs pathway. The study will provide a theoretical basis for the treatment of NASH by Shenge Formula.