基于肠道宏基因组探讨黄连-吴茱萸药对干预阿尔兹海默病的新机制

Alzheimer's disease (AD) is the most common neurodegenerative disorder in the elderly population. The pathogenesis of AD is still unknown. The latest subversive research has shown that gut microflora abnormality is closely related to the development of AD. This finding breaks the new ground in AD prevention and treatment..According to our new Chinese medicine hypothesis of AD and the new rule of "Qinggan Jieyu" in AD treatment, together with our long term research, a traditional Chinese medicine couplet medicine - “Coptis chinensis Franch - Tetradium ruticarpum” is screened as an anti-AD medicine. Our study showed this couplet medicine takes its anti-AD effect in multi targets and multi links. Meanwhile, a number of researches has confirmed Coptis chinensis Franch - Tetradium ruticarpum has a wide range of pharmacological effects on digestive system. Thus, we propose a new hypothesis that Coptis chinensis Franch - Tetradium ruticarpum may play a role in fighting AD by regulating the gut microecosystem and associated brain-gut axis..Based on this new hypothesis, this study will focus on the effects of Coptis chinensis Franch - Tetradium ruticarpum on gut microecosystem, and will further explore the anti-AD mechanism of this couplet medicine using metagenomics and bioinformatics techniques. Our study will provide scientific basis for further development and clinical application of Coptis chinensis Franch - Tetradium ruticarpum s. Meanwhile, our research will further clarify the relationship between gut microbes and Alzheimer's disease and offers new ideas for AD prevention and treatment.

阿尔茨海默病（AD）是老年人群最常见的中枢神经系统退行性疾病，目前病因未明。最新颠覆性研究成果表明：肠道微生态异常与AD发生发展关系密切，为AD防治研究开辟了全新领域。. 黄连-吴茱萸是基于“病位在脑、病本在肝”的AD中医病理新假说和“清肝解郁为主”的治疗新法则，结合多年研究优选出来的抗AD核心药对。我们多年研究表明黄连-吴茱萸药对有多环节、多靶点抗AD作用；同时大量研究工作已证实黄连-吴茱萸药对对消化系统有着广泛的药理作用。由此我们提出新假说：黄连-吴茱萸药对可能通过调控肠道微生态系统及其直接关联的脑-肠轴发挥抗AD作用。. 本项目基于这一新假说，运用宏基因组和生物信息学技术，重点观察黄连-吴茱萸对肠道微生态系统及其脑-肠轴的影响，探讨黄连-吴茱萸抗AD的全新机制，为其其进一步临床应用和研究开发提供科学依据；同时进一步阐明肠道微生物与AD的关系，为AD防治研究提供新靶点新思路。

最新研究成果表明肠道微生态异常与阿尔茨海默病（Alzheimer’s Disease, AD）发生发展关系密切。课题组多年研究表明黄连-吴茱萸药对有多环节、多靶点抗AD作用。为了进一步探究黄连-吴茱萸药对抗AD作用，项目选用5月龄的C57BL/6小鼠10只作为对照组，5月龄的APP/PS1双转基因AD模型小鼠50只，随机分为模型组、安理申组、左金丸高剂量组、左金丸中剂量组、左金丸低剂量组各10只，并分别给予不治疗、安理申治疗或不同剂量左金丸灌胃治疗12周。第13周进行水迷宫实验，实验结束后收集各组10只小鼠的新鲜粪便，进行16S rRNA扩增子测序。水迷宫实验表明，与AD模型组相比，左金丸中剂量组和安理申组在潜伏期、总路程等评价指标中的表现显著提高，且左金丸中剂量组表现最好，进一步证实了黄连-吴茱萸药对的抗AD作用。16S rRNA扩增子测序实验表明，基于Alpha多样性分析，对照组的微生物多样性最高，AD模型组的微生物多样性次之，而安理申组和左金丸不同剂量组的微生物多样性最低。根据门、纲、目、科、属和种不同分类水平的物种丰度聚类分析显示，对照组与左金丸中剂量组和安理申组距离最接近，而与AD模型组距离较远，表明左金丸中剂量组和安理申组改变了AD模型小鼠的微生物组成，使其更接近健康状态的对照组。基于Beta多样性分析，UPGMA聚类分析(Weighted Unifrac距离矩阵)的结果同样显示安理申组和左金丸中剂量组与对照组的微生物组成更接近，表明用药能够一定程度改变微生物的组成和丰度。本研究进一步论证了黄连-吴茱萸药对的抗AD作用，阐述了黄连-吴茱萸药对可能的抗AD机制，并提出黄连-吴茱萸药对通过调整AD模型小鼠的肠道微生态系统，经脑-肠轴影响AD模型小鼠，改善认知功能障碍，并提示肠道微生物是AD治疗干预可供选择的靶标。