基于“肠-肝”轴和靶向Keap1/Nrf2信号通路研究畲药三叶青干预NASH的药效物质和作用机制

非酒精性脂肪性肝炎（NASH）是非酒精性脂肪性肝病（NAFLD）发展过程中的一个病变阶段。氧化应激、脂质过氧化、炎症反应和胰岛素抵抗为其关键诱因。传统和现代医学表明畲药三叶青有显著抗炎作用，但其干预NASH的机理不清。本项目拟采用肠道炎症损伤的NASH小鼠模型，研究组织病理变化、肠道屏障功能、氧化应激、炎症因子和相关蛋白表达等内容，阐明三叶青通过“肠-肝”轴和Keap1/Nrf2信号通路干预NASH的药理活性；基于体外消化、Caco2细胞单层膜、体内代谢模型，揭示其活性成分的肠道吸收和体内代谢规律，挖掘与NASH相关的潜在药效物质；通过同源建模、分子对接和生物膜表面干涉技术靶向筛选与Keap1蛋白结合的关键药效物质；并采用Caco2和L02细胞炎症模型，验证关键药效物质通过靶向Keap1/Nrf2信号通路干预肠道炎症和NASH的分子作用机制。本课题可为三叶青防治NASH提供理论依据。

Non-alcoholic steatohepatitis (NASH) is a pathological stage in the development of non-alcoholic fatty liver disease (NAFLD). Oxidative stress, lipid peroxidation, inflammation and insulin resistance are key factors in the development of NASH. Traditional and modern medicine show that Tetrastigma hemsleyanum has significant anti-inflammatory effects, but the mechanism of its intervention in NASH is unclear. This project intends to elucidate the pharmacological activities of the intervention of Tetrastigma hemsleyanum in NASH via “Gut-Liver” axis and Keap1/Nrf2 signaling pathway by using a NASH model with intestinal inflammation and evaluating histopathological changes, intestinal barrier function, oxidative stress, inflammatory factors and related protein expression; To reveal intestinal absorption and internal metabolism of its bioactives based on in vitro digestion, Caco2 monolayer and in vivo metabolism models, and identify the potential pharmacodynamic components related to NASH; To further screen the key pharmacodynamic components that binds to Keap1 protein by use homology modeling, molecular docking and biological membrane surface layer interference (BLI) technology; To prove the molecular mechanism of the intervention of key pharmacodynamic components in intestinal inflammation and NASH by employing Caco2 and L02 cell models. It provides theoretical basis for the further prevention and treatment of NASH by Tetrastigma hemsleyanum.