从“NLRP3/IL-1β”信号通路探究心理应激在NASH发病中的作用及四逆散的干预机制

The incidence of nonalcoholic fatty liver disease (NAFLD) increases year by year, and when it develops to nonalcoholic steatohepatitis (NASH), it would like to be dangerous. In recent years, chronic psychological stress has induced and aggravated the happening of NAFLD , is worthy of attention. Our research group in the preliminary study has found that chronic psychological stress not only induced NAFLD, and would promote the occurrence of NASH, and Frigid Extremities Powder ,which is the formulas of soothing the liver has a significant therapeutic effect. Based on the theory of " emotional pathogenesis" of Traditional Chinese medicine and the theory of " liver constraint transforming into fire, liver yin internal deficiency ", we focus on the relationship of " liver fire" and inflammation, and focus on the key route "NLRP3 / IL-1β pathway" The NLRP3 / IL-1β pathway is a key link in this process and may be the main target of Frigid Extremities Powder in the treatment of NASH. The NLRP3 / IL-1β pathway is a key step in the process of neurological stress. This study will first confirm the pathogenesis of chronic stress on NAFLD / NASH, and then explore its internal mechanism from the perspective of "nerve-inflammation-metabolism" network disorder, and gradually focused on the NLRP3 / IL-1β pathway by inflammatory reaction, and to explore the promotion of the pathogenesis of NASH, and finally to observe the role of liver and Frigid Extremities Powder on the pathological process and the target, to explore the rationality of the prevention and treatment of the disease. The project will complement the pathology of NAFLD / NASH in modern society and enhance its level of prevention and control. In addition, will also enrich the traditional Chinese medicine " emotional pathogenesis " and " liver constraint transforming into fire, liver yin internal deficiency " theory of modern biological understanding.

近年来，慢性心理应激（PS）诱发和加重非酒精性脂肪肝（NAFLD）尤其是非酒精性脂肪肝炎（NASH）的现象，值得高度关注。结合本课题组前期工作，基于中医“情志致病”理论和“肝郁化火、内耗肝阴”病机认识，关注“肝火”和炎症关联，聚焦炎症关键环节“NLRP3/IL-1β通路”，提出假说：PS通过“神经-炎症-代谢”网络间影响促进NASH病理进程，NLRP3/IL-1β通路是此过程的关键环节，也可能是四逆散治疗NASH的主要靶点。本项目首先确证PS对NAFLD/NASH的病理促进作用，然后从“神经-炎症-代谢”网络失调角度以及NLRP3/IL-1β通路，探究PS干预NASH的具体机制，最后观察疏肝中药四逆散的治疗效应，探讨中西医结合防治此病的合理性。本项目将补充对现代社会NAFLD/NASH的病理认识，提升防治水平。此外，还将丰富对中医“情志致病”和“肝郁化火，内耗肝阴”理论的现代生物学认识。

非酒精性脂肪肝病（NAFLD）已被公认为全球最主要的公共卫生问题之一。越来越多的研究表明，NAFLD患病不局限于高脂、高热量饮食，还涉及了环境因素和宿主遗传学之间复杂相互作用，慢性心理应激便是NAFLD可能的重要病因之一。值得一提的是，心理应激理论与中医情志致病理论具有多层面的一致性，结合“脏腑体用相关”学说理论观点，聚焦炎症关键环节“NLRP3/IL-1β通路”，提出假说：CS通过“神经-炎症-代谢”网络影响促进NASH病理进程，NLRP3/IL-1β通路是此过程的关键环节，也可能是四逆散治疗NASH的主要靶点。本项目通过网络药理学方法、整体动物实验和体外细胞实验三个部分的研究，确证了CS对NAFLD/NASH的诱发和加重作用及四逆散对于NLRP3/IL-1β等炎症因子的调节机制。首先，网络药理学方法预测慢性心理应激与NAFLD之间存在79个交互靶点，其中31个核心靶点，21个相关作用通路（6个核心作用通路）均与炎症相关，尤其聚焦于NLRP3炎症小体的激活与炎症因子释放。动物实验采用CS单独或复合高脂饮食（HFD）诱导模型，从“神经-炎症-代谢”三个层面分别观察。结果显示：相较空白组，CS组出现了脂质堆积、血清糖皮质激素（GC）含量增高及NLRP3、NF-κB、ASC、Caspase-1等炎症信号的高表达。相较HFD组，复合组亦出现了类似结果。得出结论：慢性心理应激以介导炎症反应为核心通过“神经-炎症-代谢”网络启动、加重NAFLD疾病过程。再采用网络药理学结合体内外实验验证四逆散药效，结果表明，四逆散以抑制NLRP3炎症小体及其调控的炎症因子为核心，通过“神经-炎症-代谢”网络多通路、多靶点治疗慢性心理应激相关NAFLD。本项目将补充对现代社会NAFLD/NASH的病理认识，提升防治水平。此外，还将丰富对中医“情志致病”和“肝郁化火，内耗肝阴”理论的现代生物学认识。