基于Galectin-3调控胰岛素抵抗的青钱柳黄酮治疗非酒精性脂肪肝炎物质基础研究

The main features of nonalcoholic steatohepatitis (NASH) include accumulation of lipid droplets, inflammation and apoptosis of hepatocyte, which will deteriorate to cirrhosis and even hepatocellular carcinoma. Galectin-3 is reported to be closely related to lipid metabolism, inflammation and insulin resistance. Therefore, Galectin-3 is likely to play an important role in the development of NASH. Cyclocarya paliurus is widely used in the folk, which has the activity of hypolipemic and hypoglycemic. However, there has not been a report on the study of the treatment of NASH by Cyclocarya paliurus. In our research, we have proved that total flavonoids of Cyclocarya paliurus can inhibit insulin resistance and reduce Galectin-3 level, But it's not clear which specific substance regulates insulin resistance. The mitochondrial damage of hepatocytes is an important feature of NASH under insulin resistance. In cell models, we prove that Cyclocarya paliurus flavonoids extract can enhance the stability of mitochondrial membrane, but how Galectin-3 intervenes in the process is not clear? Based on this arguments, we hypothesized that Galectin-3 mediated the oxidative damage of hepatocytes and insulin resistance in the development of NASH, and that the two pathways could be interfered with by Cyclocarya paliurus flavonoids. This project for the first time proposed the mechanism for the treatment of NASH, which is Cyclocarya paliurus flavonoids regulating insulin resistance and liver mitochondria membrane stability, based on.Galectin-3, in order to reveal the mechanism of the Cyclocarya paliurus flavonoids treatment of NASH, and to provide the scientific basis for finding NASH therapeutic targets. This method can serve as a support for the development of material basis of traditional Chinese medicine.

非酒精性脂肪肝炎（NASH）的主要特征有脂滴堆积、炎症及肝细胞凋亡，会恶化为肝硬化乃至肝癌。Galectin-3与脂代谢，炎症及胰岛素抵抗（IR）关系密切，极可能在NASH中也起重要作用。青钱柳具有降血脂，降血糖等活性，但尚无治疗NASH的研究报道。我们在大鼠上证明青钱柳黄酮能够抑制IR，降低Galectin-3水平，但调节IR的具体物质尚不清楚。IR状态下肝细胞线粒体损伤是NASH的重要特征之一，我们在细胞上证明青钱柳黄酮能够增强肝细胞线粒体膜的稳定性，但Galectin-3如何介入这一过程尚不清楚。基于此，我们假设青钱柳黄酮能干预Galectin-3介导的NASH相关的肝细胞损伤和IR。本项目首次提出了青钱柳黄酮基于Galectin-3调控IR，稳定肝细胞线粒体膜治疗NASH的机制，为揭示青钱柳黄酮治疗NASH，以及寻找NASH的治疗靶点提供科学依据，为中药的药效物质基础的研究提供思路。

非酒精性脂肪肝炎（NASH）的主要特征有脂滴堆积、炎症及肝细胞凋亡，会恶化为肝硬化乃至肝癌。Galectin-3与脂代谢，炎症及胰岛素抵抗（IR）关系密切，极可能在NASH中也起重要作用。青钱柳具有降血脂，降血糖等活性，但尚无治疗NASH的研究报道。IR状态下肝细胞线粒体损伤是NASH的重要特征之一，但Galectin-3如何介入这一过程尚不清楚。基于此，我们假设青钱柳黄酮能干预Galectin-3介导的NASH相关的肝细胞损伤和IR。.本项目系统研究了以槲皮苷、异槲皮苷、阿福豆苷等为主要活性成分的青钱柳黄酮（CPF）的定性、定量分析方法；并完成其改善胰岛素诱导HepG2细胞的IR的药效评价；以高脂高胆固醇高糖饮食诱导C57小鼠NASH模型，评价CPF改善模型动物血糖、血脂、炎症因子、肝功能、体重以及脏器指标等的作用。采用分子对接，Western Blot，RT-PCR，IHC等技术评价CPF对NASH模型动物炎症因子，线粒体功能以及Galectin-3等关键蛋白水平的影响；并利用药代动力学-药效学研究体系，分析血药-时间-效应三者的关系，结合涉及氧化应激，三羧酸循环，氨基酸代谢，脂质代谢等多维度的代谢组学分析技术平台，完成CPF改善线粒体代谢及NASH动物病理状态的特异性分析；最终基于科学的实验数据证实了Galectin-3 是连接炎症和调控胰岛素敏感性的关键分子，而 IR和细胞因子介导了肝脏炎症及纤维化导致形成 NASH，因此，调控 Galectin-3水平是 NASH 治疗的新途径。.在明确的物质基础及作用机制的前提下，揭示了青钱柳黄酮基于Galectin-3调控IR，稳定肝细胞线粒体膜治疗NASH的机制，为中药的药效物质基础的研究提供思路，守正创新，完善中药新药开发思路，为中医药现代化、国际化提供支持。