基于肠道菌群与肠黏膜免疫对肠上皮细胞通透性的交互式调控研究汤剂微粒体系促活性成分吸收机制

The early results of our research group exhibit that the particles in Chinese Medicine decoction can significantly improve the intestinal absorption of active ingredients. Based on the regulations of intestinal flora and intestinal mucosal immune system on intestinal epithelial permeability, we hypothesis that the decoction particles can influence the balance of intestinal flora and intestinal mucosal immunity and lead an interactive control of intestinal epithelial permeability, which might be the principal mechanism of its promotive effects on active ingredient absorption in intestine. This project plans to choose the Astragalus membranaceus and Berberin as the model medicine and collect the decoction particles from single or compound decoctions. Firstly, the transport/absorption approaches of decoction particle and its influences on intestinal flora composition will be investigated, to evaluate its disposal process in intestine. Secondly, the activation/inhibition of TLR-κB signaling pathway, the differentiation of immune-related cells, and the expression of related effectors in intestinal tissues will be investigated, to clarify the influence mechanisms of decoction particles on the ultrastructure of cellular tight junction between intestinal epithelial cells, as well as the expression of related proteins. Finally, the correlationship and correlation mechanism between decoction particles, intestinal mucosal immunity/intestinal flora balance, and intestinal epithelial permeability will be discussed. This project will contribute to clarify some common problems of Chinese Medicine decoction on active ingredients absorption in intestine, and will prove theoretic bases to explain the scientific implications of decoction.

前期研究发现：中药汤剂微粒可显著促进活性成分肠吸收。基于肠粘膜免疫系统和肠道菌群对肠上皮细胞通透性的调控作用，我们提出假说 “汤剂微粒通过影响肠黏膜免疫与肠道菌群的平衡状态，交互式调控肠上皮细胞通透性，是其促活性成分吸收的主要机制”。课题拟分别制备黄芪和黄连的单/复方汤剂并分离收集微粒；首先，考察汤剂微粒的肠转运/摄取途径及对肠道菌群组成的影响，确定其肠内处置过程；然后，研究汤剂微粒对肠组织TLR-κB信号通路激活/抑制状态、免疫相关细胞分化水平、相关效应分子表达水平的影响，明确微粒影响肠上皮细胞紧密连接超微结构与相关蛋白表达水平的机制；最后，探讨汤剂微粒、肠黏膜免疫/肠道菌群平衡、肠上皮细胞通透性三者间相关性与关联机制。课题的实施有助于阐明中药汤剂促活性成分吸收的共性机制，为诠释中药汤剂的科学内涵提供理论基础。

汤剂是中医传统给药剂型，基于肠道菌群与肠黏膜免疫系统对肠上皮细胞通透性的交互式调控作用，本项目提出“中药汤剂微粒通过其多糖成分调节肠黏膜免疫与肠道菌群的平衡状态，使肠上皮细胞与肠黏膜免疫细胞分泌具有不同生物学功能的效应分子，调节肠上皮细胞紧密连接相关蛋白表达水平和组装，实现对肠上皮细胞通透性的交互式调控，从而改善中药活性成分肠吸收特性。研究结果发现：植物源中药材水煎煮过程中多糖大分子溶出并自组装形成微纳米尺寸微粒，汤剂多糖微粒通过调节小肠段菌群分布与肠黏膜淋巴细胞活化/分化，影响肠上皮细胞紧密连接蛋白表达水平和组装，实现对肠上皮细胞通透性的动态调控，从而提高活性成分肠吸收。本项目创新性的从汤剂多相分散体系与肠道微生态两个复杂系统的交互作用角度探讨汤剂有利于活性成分肠吸收的作用机制。这一研究思路不受活性成分自身理化性质与生物活性的限制，对“汤剂”这一中药经典剂型具有普适性价值，而且拓展了中药制剂生物药剂学研究的视角。本项目迄今为止已发表学术论文14篇，其中SCI期刊论文7篇（中科院一区期刊论文3篇、二区期刊论文4篇）；在本项目资助下已有4名硕士研究生顺利完成学习并获得学位；以本项目为基础，主持人继续深入中药汤剂科学内涵研究，获批2019年安徽省自然科学基金杰青项目与2022年安徽省教育厅高校自然科学研究重大项目。