从下丘脑弓状核POMC/CART通路与肠道菌群变化相关性研究肝郁脾虚证生物学基础

Liver-depression and spleen-deficiency syndrome is the chief syndrome of sub-health, and the frequent syndromes of various clinical diseases. At present, experimental studies emphasis on emotional disorders and behaviour changes relating with the liver depression, while lack of studies on spleen-stomach malfunction manifested with food-intake and body-weight decreased, and loose stool, et al...Our early study results showed POMC/CART pathway related appetites in hypothalamic arcuate nucleus(ARC) is the critical neuroendocrine basis of spleen and stomach malfunction with stress, and intestinal flora disorder lead to digestion and absorption malfunction, belongs to the external critical factors. The correlation between POMC/CART pathway and intestinal flora disorder, related spleen-stomach malfunction with liver-depression and spleen-deficiency syndrome, is our further research target..Based on the early study and literature review, we think the brain-gut-microbiota pathway is the biological basis of liver-depression and spleen-deficiency syndrome with chronic stress. So we will observe the changes of the intestinal flora and NPY/AgRP and POMC/CART neuropeptides related appetite in hypothalamus ARC, especially the relations between the intestinal flora and Leptin-ob-R-POMC/CART nervous pathway, to further more explore the biological basis of spleen and stomach malfunction in liver-depression and spleen-deficiency syndrome, and clarify the main functional mechanism and target of Xiaoyaosan (Formularies of the Bureau of People's Welfare Pharmacies) for smoothing liver to relieve depression for invigorating spleen in order to regulate appetite.

肝郁脾虚证是亚健康人群首要证候，也是临床多种疾病常见证候之一。目前实验重点集中于动物模型出现情绪障碍和行为改变等肝郁样表现的基础研究，而肝郁脾虚证机体出现脾胃运化功能失常的机制研究涉及不足。.课题组前期研究结果认为：下丘脑弓状核POMC/CART通路是应激状态脾胃失常的中枢关键环节；肠道菌群紊乱导致消化吸收功能障碍，则属其外周重要因素。二者相关性研究及与肝郁脾虚证机体出现食少、形瘦、便溏等表现的内在机制，是本课题组亟待研究解决的问题。.文献结合前期研究，本课题组提出，脑-肠-微生物途径可能为慢性应激肝郁致脾虚的内在生物学基础。所以，基于脑-肠-微生物途径，从下丘脑弓状核摄食相关通路和肠道微生物菌群变化的相关性，尤其Leptin-ob-R-POMC/CART通路与肠道特定微生物相关性变化，深入研究肝郁脾虚证脾胃运化功能失常的病理机制，及逍遥散（《局方》）疏肝健脾以调食欲的主要作用机理与靶点。

肝郁脾虚证是亚健康人群首要证候，也是临床多种疾病常见证候之一。基础研究多从神经、内分泌、免疫、消化、循环及代谢系统等方面，采用分子生物学、代谢组学、基因组学等方法，对动物模型出现情绪障碍或行为改变等肝郁样表现，或对消化功能异常等脾虚样表现的病理机制进行研究，但肝郁如何导致脾虚，由情绪障碍逐渐出现饮食减少、体重减轻、大便溏泄等脾胃运化功能失常的中间环节机制涉及研究尚不足。.本研究以每天3h连续21天捆绑束缚方式可制备慢性应激肝郁脾虚大鼠模型，模型动物表现血清瘦素水平升高，下丘脑ARC中促食欲因子NPY、AgRP表达下降，抑食欲因子POMC、CART表达增加，逍遥散具有明显调节作用；并且通过RNA干扰技术重点观测到模型动物下丘脑ARC中ob-R-CART/ POMC（α-MSH）通路可作为慢性应激肝郁脾虚动物模型脾胃运化功能失常的中枢神经内分泌基础之一，粪样小分子代谢物短链脂肪酸（Acetate、Butyrate、Proionate）和糖类（Glucose、Maltose、Ribose、Xylose）、肠道微生物（Firmicutes、Patescibacteria、Proteobacteria，Gammaproteobacteria、Saccharimonadia，Enterobacteriales，Aerococcaceae、Bacteroidaceae、Enterobacteriaceae、Prevotellaceae、Rikenellaceae、Ruminococcaceae， Adlercreutzia、Aerococcus、Bacteroides、Escherichia-Shigella、Rikenellaceae_RC9_gut_group、Ruminococcaceae_NK4A214_group、[Eubacterium]_ventriosum_group）结构与功能变化与慢性心理应激肝郁致脾虚和逍遥散作用机制相关。提示脑-肠-微生物途径可能为慢性应激肝郁致脾虚的内在生物学基础，丰富了中医肝郁脾虚证的科学内涵，阐释了逍遥散疏肝以健脾调节脾胃功能的作用机理，为其临床应用提供实验基础。