基于压力应激-肠道微生态-中枢神经炎症系统调节的开心散抗抑郁作用机制研究

As a classical TCM formula for treating major depression disorder, the clinical efficacy of Kai-Xin-San (KXS) had been confirmed and the mechasim was revealed by monitoring neurotransmitters and potentiating neurotrophic factors. Now, it has been found that disturbance of intestinal microecology should be included in pathology of depression. In details, stress induces changes of intestinal microbiota and injury of intestinal barrier, which leads to neuronal inflammaiton of central nervous system and excitation of HPA axis. Under this circumstance, depression is caused. Here, the anti-depression machanism of KXS will be further elucidated on depression animal and cell models via this newly discovered pathological mechanism. Firstly, the effect of KXS on stress system will be explored by determining the expressions of stress hormones and related receptors. Secondly, via technologies of highflux gene sequencing and ussing chamber, the effect of KXS on intetinal microbiota composition and intestinal barrier function will be elucidated. Thirdly, the effect of KXS on neuronal inflammaiton of central nervous system will be explored by determining the expressions of inflammantory factors, neurotranismitters, neurotrophic factors and related monitoring enzyemes. Finally, the action machanism of KXS on these revealed targets will be further explored on cell models. Based on these researches, we hope to elucide the anti-depression mechansim of KXS via monitoring stress-intestinal microecology-central neuronal inflammation circuit, which will be helpful for supplying more scientific evidences for development of anti-depressant.

开心散是抗抑郁经典方剂，前人研究已证实其明确的抗抑郁效用。申请者发现其不仅可调控神经递质、还可调控并增益神经营养因子发挥抗抑郁作用。近年来，国内外研究证实，抑郁症与肠道微生态紊乱密切相关。压力应激导致肠道菌群变化，损伤肠道屏障，诱发中枢神经炎症，加剧压力应激，构成系统回路，加剧抑郁程度。我们拟基于此回路，构建动物与细胞模型，深入探索开心散抗抑郁机制，拟开展以下工作：（1）通过压力激素与相应受体表达测定，探索开心散调控压力应激系统的效用机制；（2）采用高通量基因测序与Ussing Chamber等技术，阐述开心散对肠道菌群结构与肠道屏障功能的调控机制；（3）通过炎性因子、神经递质、神经营养因子与相关调控酶测定，揭示开心散调控中枢神经炎症系统的效用机制；（4）构建细胞模型深入机制研究。从而系统阐释开心散调节压力应激-肠道微生态-中枢神经炎症系统抗抑郁的作用机制，为开发抑郁症治疗药物提供科学依据

本项目在四年的研究周期内，针对开心散抗抑郁作用与压力应激-肠道微生态-中枢神经炎症系统调控，开展了如下研究，取得了阶段性成果，介绍如下。. 首先，我们构建了能够较好的模拟抑郁症病人快感缺失等特征症状的慢性应激压力小鼠抑郁症模型，结合开心散用于“心脾两虚，痰湿中阻”的证候特点，建立了符合“心脾两虚，痰湿中阻”证候特点的抑郁症病症结合动物模型。不同配伍比例的开心散给药后均有显著的减轻动物抑郁样行为的作用，其中以比例D-652（人参:远志:石菖蒲:茯苓为3:2:2:3）作用趋势最强。. 其次，我们利用16sDNA技术检测肠道菌群改变，发现压力应激显著上调肠道革兰氏阴性菌的丰度，下调革兰氏阳性菌的丰度，提示压力应激引起肠道菌群组成紊乱。同时压力应激显著破坏了肠道屏障，下调肠道屏障蛋白表达，促进炎性因子白介素-1,6与肿瘤坏死因子表达增多。模型动物脑中的炎症因子表达也显著增高，血脑屏障蛋白下调显著。此外模型动物血清与器官中的压力激素表达也显著增高，体现存在肠道炎症-压力应激-中枢神经炎症的通路。上述趋势均可以被开心散给药逆转。进一步研究发现，开心散的上述作用可以被抗生素所扭转，充分证明了脑肠轴调控是开心散抗抑郁作用的重要环节。. 最后，基于各调控环节，建立了代表性细胞模型进行效应成分筛选，获知了功效成分。将功效成分进行配伍，形成的开心散组分配伍，抗抑郁作用与开心散原方作用相当。. 项目发表相关文章7篇，SCI文章5篇，中文核心期刊2篇。申请专利1项。初步阐明了开心散调节压力应激-肠道微生态-中枢神经炎症系统抗抑郁的整合效应机制，为探索基于压力应激-肠道微生态-中枢神经炎症系统的抑郁症临床治疗策略及创新药物的开发提供了科学依据。