护肝清脂片防治非酒精性脂肪肝的信号传导网络机制研究

Nowadays, nonalcohlic fatty liver disease(NAFLD) has become a global public health problem, without interpreted nosogenesis and desirable treatment. It is currently considerd that multiple factors based on insulin resistance-oxidative stress may participate in the development of NAFLD, but the mechanisms of its intracellular signal transduction network is still unkown. On the basis of prevenient research, we presumed that Huganqingzhi tablet could improve insulin resistance, alleviate injuries of oxidative stress and the production of inflammatory factors, finally prevent the development of NAFLD ,probably by the regulation of its relevant signal transducion network. This study plans to explore the mechanism of Huganqingzhi tablet by combined experiments in vivo and vitro. At first, we will establish the rats and cell models of NAFLD. Secondly,we probe the discrepant expression spectrum of intracellular phyosphorylated protein,signal transducin, by the method of proteomics and observe their pathological changes, insulin resistance, oxidative stress, inflammatory factors and so on in NAFLD rat models. Finally, we will analyze their inherent connections bewteen the pathologic processes of NAFLD and intracellular signal transducin by bioinformatics to identify the characteristic of signal transduation network in NAFLD and represent the siganl regulation of Huganqingzhi tablet in the prevention of NAFLD, which would provide scientific basis for its application and extension in clinical.

非酒精性脂肪肝（NAFLD）已成为全球性的公共健康问题，迄今为止其发病机制尚未阐明，亦没有理想的治疗药物。目前认为胰岛素抵抗-氧化应激损伤为基础的多种因素参与了NAFLD的发病过程，但其细胞内信号传导机制仍未清楚。依据前期研究，我们推测护肝清脂片可能是通过调节细胞内有关的信号传导网络，以改善胰岛素抵抗、减轻氧化应激损伤、减少炎症因子生成，从而防治NAFLD。本项目拟采用体内与体外实验相结合的方法探讨护肝清脂片的作用机制，首先建立NAFLD大鼠模型和细胞膜型，其次应用蛋白组学方法探查细胞内的差异磷酸化蛋白表达谱-即信号传递蛋白，同时观测NAFLD病理学变化、胰岛素抵抗、氧化应激损伤、炎症因子等有关指标，最后应用生物信息学分析NAFLD病理过程与细胞内信号传递蛋白的内在联系，以明确NAFLD信号传导网络变化特征，同时阐明护肝清脂片防治NAFLD的信号传导调控机制，为其临床推广应用提供科学依据。

非酒精性脂肪肝 (NAFLD) 已成为全球性的公共健康问题，迄今为止，NAFLD的发病机制尚未阐明，亦没有理想的治疗药物。依据前期研究，我们推测护肝清脂片可能是通过调节细胞内有关的信号传导网络，以改善胰岛素抵抗、减轻氧化应激损伤、减少炎症因子生成，从而防止NAFLD。本项目拟采用体内体外实验相结合的方法探讨护肝清脂片的作用机制，建立体内体外NAFLD模型，运用iTRAQ蛋白组学技术寻找相关差异蛋白，同时观测NAFLD病理学变化、氧化应激损伤、炎症因子等有关指标，最后结合对差异蛋白的生物信息学分析结果共同地明确NAFLD信号传导网络变化特征，同时阐明护肝清脂片防治NAFLD的相关药理作用机制及可能的潜在作用靶点。. 体内体外实验结果显示护肝清脂片能显著地改善NAFLD的相关生化指标，iTRAQ蛋白定量分析结果显示NAFLD的作用机制复杂，其主要与氧化应激，细胞凋亡坏死、内质网应激、肠道菌群的代谢活动、胆汁酸代谢等有关。而护肝清脂片可以通过相关靶点如CANX、Hsp27、ACAT1、ILK、PHYH、ACSL1、FABP4、ORM、Sult2a1和ASS1等起到防治NAFLD的作用。