基于“微生物-肠-脑轴”研究石斛合剂防治糖尿病脑病的作用机制

The microbiota-gut-brain axis plays an important role in the pathological process of diabetic encephalopathy. Previous study has shown that Dendrobium mixture which was based on ziyin yiqi huoxue method, could also alleviate cognitive impairment of diabetic encephalopathy, while effectivelly treated diabetes by the regulation of glucagon-like peptide-1 (GLP-1) signal pathway. Because of the close relationship between GLP-1 secretion and gut microbiota, we speculate that the overall mechanism of dendrobium mixture on DE prevention and treatment may attribute to microbiota-gut-brain axis adjustment and insulin resistance alleviation. This project is based on the overall view of traditional Chinese medicine characteristics, take microbiota-gut-brain as the breakthrough point, using the method of pharmacodynamics and molecular biology to observe effects of dendrobium mixture on sugar and lipid metabolism, cognitive dysfunction, brain tissue pathological changes in rats DE models, as well as the gut microbiota by intestinal microecology Macro genome sequencing and Real-time PCR method. We are going to find the functional intestinal bacteria through correlation analysis method and to explore the mechanism of the ziyin yiqi huoxue method of DE prevention and treatment effects. At the end, this project will be developed into one of the modern researches of traditional Chinese medicine, and will be theoretical basis of effective prevention and treatment of diabetic encephalopathy. At the same time, we are going to make the modern science interpretation for the prevention and treatment of diabetic encephalopathy by ziyin yiqi huoxue such a traditional Chinese medicine treatment method.

微生物-肠-脑轴在糖尿病脑病（DE）的病理过程中发挥重要作用。前期研究发现,以滋阴益气活血法立方的石斛合剂可以通过调控胰高血糖素样多肽-1(GLP-1)信号通路干预2型糖尿病，同时还可以改善DE的认知障碍。由于GLP-1的分泌与肠道微生物关系密切，我们推测：通过调控微生物－肠－脑轴，改善胰岛素抵抗,是石斛合剂防治DE整体效应的重要机理。本项目依据中医药整体观的作用特点，以微生物－肠－脑轴为切入点，运用药效学、分子生物学等方法观察石斛合剂对DE动物模型糖脂代谢紊乱、认知功能障碍、脑内组织病理结构变化的影响，并结合肠道微生态学宏基因组测序和荧光定量PCR技术,观察石斛合剂对肠道菌群结构的影响，通过关联分析找到石斛合剂影响的肠道功能菌，探索滋阴益气活血法方药防治DE的作用机制。本研究将为开发疗效确切的防治DE的创新药物奠定基础，同时对滋阴益气活血法治疗消渴呆病的中医传统治法进行现代科学阐释。

本研究明确了石斛合剂可改善糖尿病认知障碍大鼠的学习和记忆能力，并且对糖尿大鼠血脑屏障有保护作用，提高紧密连接蛋白Occludin、ZO-1表达，减少炎症因子IL-6、TNF-α、LPS的表达，缓解神经炎症的进展。进一步研究发现石斛合剂改善神经炎症的作用可能与调节肠道菌群，提高结肠紧密连接蛋白Occludin、ZO-1表达而保护肠道屏障作用，改善肠-脑轴的炎症信号有关。本研究还建立了一种新型的3D培养的神经炎症细胞模型，为课题的进一步深入研究奠定了基础。通过本项目的研究，可为后续中药复方的现代研究和DE的有效防治提供方法借鉴和理论依据。