非酒精性脂肪肝是一种常见肝脏疾病，其典型病理变化是肝细胞内过多甘油三酯沉积，并出现脂滴。目前遗传因素导致的NAFLD逐渐引人关注，其中PNPLA3 rs738409基因多态性经实验证实与肝内脂肪代谢密切相关。本项目拟通过制备PNPLA3-WT及其突变体PNPLA3-I148M基因或si-PNPLA3的腺病毒表达系统，感染原代肝细胞或建立稳转的Huh-7细胞，检测各组肝脏细胞中脂滴代谢相关因素的差异，分析PNPLA3与脂滴蛋白OXPAT的相互作用，探究PNPLA3在脂滴脂肪代谢中的作用，以及PNPLA3-I148M影响脂滴脂肪蓄积的分子机制。体内实验通过建立NAFLD大鼠模型，腺病毒介导PNPLA3-WT及PNPLA3-I148M基因在肝脏中过表达或沉默表达，检测血清及肝脏中各脂肪代谢指标，分析肝脏脂肪病变情况，体内验证PNPLA3-WT及PNPLA3-I148M与NAFLD发生发展的关系。

Nonalcoholic fatty liver disease (NAFLD) is a common disease, and its hereditary factors have attracted more and more attention. The typical pathological feature of liver steatosis is the lipid deposition in liver cells in the form of lipid droplets (LDs).In recent years, PNPLA3 polymorphism has become a hot topic in research of NAFLD, and associated with liver inflammation development and disease progression. To examine the role of PNPLA3 in triglyceride metabolism and elucidate the mechanism by which the I148M substitution affects liver lipid content, we delivered human PNPLA3 cDNA, PNPLA3-I148M and siRNA-PNPLA3 in adenovirus vector, which are transferred to primary liver cells or Huh-7 cells. Then the factor of lipid droplets metabolism were detected in each cell group. We also indentify the PNPLA3-interacting protein-OXPAT and facilitate the illustration of the molecular mechanism of PNPLA3. Adenoviral vectors are transferred to rat model of NAFLD. Then the blood biochemical parameters and lipid metabolism of liver were measured, and the hepatic steatosis was evaluated by immunohistochemical analysis to verify the relationship between the PNPLA3-I148M and development of NAFLD. The research explores the molecular mechanisms and provides strong theoretical basis and technical support of clinical treatment to NAFLD.