葛根芩连汤对NASH肠-肝对话免疫模式识别中NLRP3调控机制研究

The studies supported by the Innovation team of Beijing University of Chinese Medicine (No. 2011-CXTD-24), Wang Bao-en liver fibrosis research fund (No.2013-xjs) and Self-selected subject of Beijing University of Chinese Medicine (No.2013-JYBZZ-XS-153) discovered that: Gegenqinlian Decoction treatment is a potential way to manage NASH via regulating lipid and glucose metabolism and suppressing inflammatory factors, such as IL-1、IL-10、TNF-α. The regulatory mechanism research on NLRP3 in immune pattern recognition in NASH by Gegenqinlian Decoction based on gut-liver axis will be conducted by using NASH rat model established with high fat diet, free fatty acid-induced HepG2 cells and intestinal Caco-2 cells induced under high-fat environments. We take a lead research on molecular mechanism of NLRP3 in immune pattern recognition in NASH based on gut-liver axis, and make a detail research on the expression of ASC, pro-caspase-1(P10), SGT1, HSP90 in NASH in vivo and in vitro, which would confirm the acting targets of NASH by Gegenqinlian Decoction, uncovering the regulatory mechanism of Gegenqinlian Decoction on NLRP3 based on gut-liver axis. What’s more, the difference in regulating NLRP3 will be explicated by comparing their differences between Gegenqinlian Decoction and Glutamine, which would make a great contribution to clinical using of Gegenqinlian Decoction eventually.

北京中医药大学创新团队项目等研究表明：葛根芩连汤能够通过改善糖脂代谢紊乱及炎症因子IL-1、IL-10、TNF-α发挥防治非酒精性脂肪性肝炎（NASH）的作用。采用高脂饲料复制大鼠NASH 模型，结合游离脂肪酸诱导HepG2细胞建立的NASH体外模型以及高油脂环境下的肠Caco-2细胞体外模型，研究葛根芩连汤对NASH肠-肝对话免疫模式识别中NLRP3调控的分子机制。率先进行NASH 肠-肝对话免疫模式识别中NLRP3调控机制的研究，重点研究NASH 体内外模型中ASC 、pro-caspase-1（P10）、SGT1、HSP90基因转录及蛋白表达的相关性，阐明葛根芩连汤干预NASH的作用靶点，揭示葛根芩连汤对NASH肠-肝对话免疫模式识别中NLRP3的调控机制；比较葛根芩连汤与阳性药谷氨酰胺干预NASH的疗效差异，明确二者调控NLRP3的作用差异点，为葛根芩连汤的临床应用提供科学依据。

本课题研究了葛根芩连汤对非酒精性脂肪性肝炎肠-肝对话免疫模式识别中NLRP3 调控的分子机制。研究发现：高脂饲料饲喂的大鼠，小肠中炎症活化趋势强，炎症小体NLRP3中激活因素与稳定因素的平衡被打破，以炎症小体激活因素活化为主；肝脏中炎症小体亦有活化，但较易形成“高位”新的平衡；葛根芩连汤干预可减低激活因素活化程度、提高稳定因素活化程度，利于炎症小体内部平衡的恢复。同时，葛根芩连汤可以明显降低高脂饲料诱导的NASH大鼠模型血清sIgA的水平，增加肠道紧密连接蛋白ZO-1表达并降低肠道PGE2蛋白水平，达到调节肠道屏障的功能，从而对NASH的防治起到积极作用。另外，葛根芩连汤能够改善对高脂饮食诱导的NASH大鼠模型肠道菌群分布，尤其是低剂量组，对于Firmicutes, Clostridia, Lactobacillus, bacilli 及Erysipelotrichales等菌群丰度有一定提升，与正常组肠道菌群分布较为相似。该研究为葛根芩连汤应用防治非酒精性脂肪性肝炎提供了更多的科学依据。相关研究发表文章5篇，其中中文核心期刊2篇，SCI文章3篇。