应用宏基因组学探讨抑郁症肝郁乘脾的发生机制

The wood over-restraining the earth, the important theory about the five elements theory of traditional Chinese medicine (TCM), has yet to require further elucidation for its scientific connotation. Previous studies have presented evidence that gut microbiota has play an important role in the occurrence of depression. Liver depression over-restraining the spleen, which is the same as the wood over-restraining the earth, is the important pathogenesis and common syndrome in TCM. A previous investigation by us had found that remarkable changes took place in the construction of gut microbiota among the depression patients with liver depression over-restraining the spleen, and it could be recover by the treatment of Xiaoyao Pill. Accordingly, we theorize that the abnormal gut microbiota may play an important role in the occurrence of liver depression over-restraining the spleen. In this project, we aim to investigate the change characteristics of gut microbiota with liver depression over-restraining the spleen in depression patients clinically, and in the making animal model of the transgenic mouse with depression, by adopting the method of disease integrating with syndrome, the intervention means of Xiaoyao Pill, and the technology of metagenomic; eventually, conducting bioinformatics method for integrated analysis, we will ascertain how the unbalance of homeostasis in gut microbiota works in the development of depression with liver depression over-restraining the spleen, reveal the scientific connotation of the wood over-restraining the earth, and confirm the regulation mechanism of gut microbiota in the treatment of depression by Xiaoyao Pill, intending to provide scientific evidence for clinical adoption.

“木乘土”是中医五行学说的重要理论，但其科学内涵尚未阐明。诸多证据表明：肠道微生物在抑郁症发病中起到重要作用。中医学认为肝郁乘脾（木乘土）是抑郁症的重要病机和常见证候。我们前期研究发现抑郁症肝郁乘脾患者肠道菌群结构发生显著的变化，而给予逍遥丸治疗后可促进其肠道菌群异常的恢复。因此推测肠道菌群的稳态失衡可能促进了肝郁乘脾的发生发展过程。本项目拟采用病证结合方法，以逍遥丸作为干预手段，应用宏基因组学方法，首先在临床上探讨抑郁症肝郁乘脾患者肠道菌群的变化特征，其次利用抑郁症转基因小鼠制作动物模型，探讨小鼠模型肠道菌群的变化特征；再次，应用粪菌移植技术，利用无菌小鼠探讨肠道菌群对肝郁乘脾发生的影响。最后，利用生物信息学方法进行整合分析，阐明肠道菌群的稳态失衡在抑郁症肝郁乘脾发生中的作用，从肠道微生物角度揭示中医“木乘土”理论的科学内涵，并明确逍遥丸对肠道微生物的调节机制，为其临床应用提供科学依据。

“木乘土”是中医五行学说的重要理论，但其科学内涵尚未阐明。我们推测肠道菌群的稳态失衡可能促进了肝郁乘脾的发生发展过程。为阐明加味逍遥颗粒治疗抑郁症的肠道菌群调节机制，本项目采用病证结合方法，以加味逍遥颗粒作为干预手段，应用16S区域扩增子测序分析方法，初步分析发现：与正常人比较，抑郁症肝郁脾虚患者肠道菌群结构有明显差异，差异主要集中在放线菌、拟杆菌及厚壁菌；其次，成功制作抑郁症肝郁乘脾小鼠模型，开展加味逍遥颗粒干预抑郁症肝郁乘脾的动物实验研究，结果发现加味逍遥颗粒组在进食量、体重等指标方面，在糖水偏好、强迫游泳、自发活动、Y-迷宫、明暗箱穿梭等行为学实验方面显著改善抑郁小鼠的状态，其作用与西药舍曲林相当；再者，本项目采取口服抗生素方法模拟无菌小鼠，并在给予中剂量加味逍遥颗粒干预后行为学表现均优于模型组，其作用亦与西药舍曲林相当。目前动物实验所有血液及组织标本已送出检测，等待检测结果，后续将对结果进行整理分析，撰写SCI论文及发表，将进一步阐明肠道菌群在抑郁症肝郁乘脾发生中的作用，揭示加味逍遥颗粒治疗抑郁症的肠道菌群调节机制，从肠道微生物角度揭示中医五行学说――“木乘土”理论的科学内涵。本项目目前已经发表中文学术论文5篇，其中中文核心2篇，统计源3篇；项目负责人在执行周期内招收硕士研究生1名。