5-HT3R与CB1R协同介导情绪调控脑区GABA释放在PMDD肝气逆郁两证中的作用

Premenstrual dysphoric disorder (PMDD), the serious type of premenstrual syndrome(PMS), is a common disease with great harms and complicated pathogenesis. It has been the research hots how to clinically sub type PMDD for effectiver prevention and treatment. Previously we found out Gan-qi invasion and Gan-qi depression were two main syndromes of PMS resulted from malfunction of Gan. Furthermore, there were abnormal changes in levels of 5-hydroxytryptamine (5-HT), γ-amino-butyric acid (GABA), 5-HT3R gene and GABAR gene in brains of these two syndrome of PMS.Some of these abnormal changes were same while the others were different. It has been known that 5-HT3R and cannabinoid 1 receptor (CB1R) coexpressed in presynaptic membrane of GABAergic neurons in brain regions regulating mood respectively mediating promotion and inhibition of GAGA release. There are no reports of 5-HT3R and CB1R in PMDD. From these we put forward the hypothesis that 5-HT3R and CB1R comediate release of GAGA in mood regulating brain regions respectively play different roles in pathogenesis of PMDD with Gan-qi invasion and Gan-qi depression. In the study, we will detect location, expression levels and binding rates with the ligands of 5-HT3R and CB1R in brain regions of model rats.The specific agonist and antagonist of 5-HT3R and CB1R will all be used to investigate their different roles in comediating release of GAGA in pathogenesis of PMDD with Gan-qi invasion and Gan-qi depression. The study will provide scientific basis for PMDD subtype research and therapy as well as provide carrier,targets and experimental basis for exploring brain micromechanism of Gan in affective regulations.

经前烦躁障碍（PMDD）为重度经前期综合征（PMS），危害严重，病机复杂不明，其亚型区分和有效防治是当今研究热点。我们前期发现，肝疏泄失常所致肝气逆证、郁证为PMS主证，均存在脑内5-HT、GABA含量和5-HT3R、GABAAR基因表达异常，改变呈同中存异。已知5-HT3R与大麻素受体CB1R共表达于情绪调控脑区GABA能神经元，分别介导促进、抑制GABA释放，未见PMDD相关报道。故推测：5-HT3R与CB1R协同介导情绪调控脑区GABA释放共同参与PMDD肝气逆郁两证，且作用不同。本项目拟利用病证结合大鼠模型，检测不同脑区5-HT3R、CB1R的分布表达及其配体结合率，运用特异性激动、拮抗剂检测其协同介导GABA释放在中医肝疏泄失常所致PMDD肝气逆郁两证中的作用区别及微观机制，为PMDD亚型区分与防治提供科学依据；为探索肝主疏泄调畅情志脑中枢机制提供研究载体、具体靶向和实验依据。

经前烦躁障碍（PMDD）为重度经前期综合征（PMS），危害严重，病机复杂不明，其亚型区分和有效防治是当今研究热点。本研究主要研究5-HT3R与CB1R协同介导情绪调控脑区GABA释放共同参与PMDD肝气逆郁两证，且作用不同。本项目通过制备病证结合动物模型，综合应用行为药理、ELISA（酶联免疫法）、免疫荧光、Western blot、RT-qPCR等多种实验手段，利用病证结合大鼠模型，检测不同脑区5-HT3R、CB1R的分布表达；运用特异性激动、拮抗剂检测其协同介导GABA释放在中医肝疏泄失常所致PMDD肝气逆郁两证中的作用区别及微观机制。最终揭示5-HT3R和CB1R协同介导GABA释放在PMDD 肝气逆、郁两证中的不同作用机制，明确中药相应作用靶点；揭示参与PMDD肝气逆、郁两证发病机制中众多影响因素之间的相互关系。我们认为，5-HT3R与CB1R共同介导参与PMDD肝气逆、郁两证发病机制中，可以单独和共同调节GABA的含量，为PMDD亚型区分与防治提供科学依据，使肝疏泄失常的微观机制得到阐明成为可能，创新中医学肝调畅情志理论。