GABABR介导GABA调节中缝背核5-HT能神经元在愤怒郁怒中的作用与机制研究

Anger is one of the common negative emotions harmful to human health. It contains unclear to the mechanism of anger inducing many disorders and diseases. In the Chinese traditional medicine theories, anger induces diseases by hurting Gan-one of functional systems in the body. In our previous studies, we found out that abnormal functions of Gan would result different expressing patterns of anger. Gan excessively working would result in anger-out, while it did not work enough resulting in anger-in. The pathologies of anger-our and anger-in both related to 5-hydroxytrytamine (5-HT) and γ-aminobutyric acid (GABA) in the brain. The changes of 5-HT and GABA were obviously different in brains of anger-out and anger-in rats. Furthermore, the expression levels of GABA receptor B (GABABR) were also markedly different in brains of those model rats. Our results suggest that GABABR inducing GABA effects play important roles in anger-out and anger-in pathologies. It is well known that 5-HT in the brain is mainly synthesized in serotonin neurons of dorsal raphe neucleus and then is delivered to other brain regions by neural fibers. It was confirmed that serotonin neurons in dorsal raphe neucleus were regulated by GABA. There is no report about anger and GABA modulating serotonin neurons in dorsal raphe neucleus. In the present project, we will demonstrate the roles and mechanism of GABABR modulated serotonin neurons in dorsal raphe neucleus of anger-out and anger-in male rats with biological techniques in order to define the targets of some regulating-Gan herbal formula and to provide new data to explain the theory of Gan regulating emotions in the Chine traditional medicine.

怒是影响人类健康的重要情志因素，其致病机制尚未完全明确。"怒伤肝"是中医经典理论，本质有待于进一步揭示。我们前期发现怒有愤怒和郁怒两种表现形式，分别由肝疏泄太过与不及所致，病机均与脑内5-HT、GABA有关，但存在明显差异，且两者模型大鼠多个脑区GABA受体GABABR表达显著不同，提示GABABR 介导的GAGA效应在愤怒、郁怒中起重要作用。已知脑内5-HT主要来自中缝背核5-HT能神经元，经纤维投射至不同脑区；该类神经元受GABA调节，但与怒的相关研究国内外未见报道。本项目拟在已有工作基础上，利用愤怒郁怒大鼠模型，借助于脑立体定位显微注射、脑微透析、免疫电镜双标等现代生物学技术，深入探索GAGABR介导GAGA效应对中缝背核5-HT能神经元释放5-HT调节前额区、下丘脑、海马功能的影响在愤怒、郁怒中的作用及机制，明确调肝方药相应作用靶点，为揭示"肝主疏泄调畅情志"本质提供新的科学依据。

怒是影响人类健康的重要情志因素，其致病机制尚未完全明确。“怒伤肝”是中医经典理论，本质有待于进一步揭示。我们前期发现怒有愤怒和郁怒两种表现形式，分别由肝疏泄太过与不及所致，病机均与脑内5-HT、GABA有关，但存在明显差异，且两者模型大鼠多个脑区GABA受体GABABR表达显著不同，提示GABABR介导的GAGA效应在愤怒、郁怒中起重要作用。已知脑内5-HT主要来自中缝背核5-HT能神经元，经纤维投射至不同脑区；该类神经元受GABA调节，但与怒的相关研究国内外未见报道。本项目在已有工作基础上，利用愤怒、郁怒大鼠模型，借助于脑立体定位显微注射、ELISA、免疫电镜双重标记、免疫荧光双重标记等现代生物学技术，逐步开展了系统、深入的研究工作。项目不仅进一步完善了愤怒、郁怒大鼠模型制备与评价方案，申请了国家发明专利，而且首次发现愤怒、郁怒作为怒的两种表现形式，均可导致中缝背核GAGA含量和GABABR1表达异常升高，进而GABABR1介导GABA抑制中缝背核5-HT能神经元产生5-HT，导致关键脑区（下丘脑、前额区、海马）5-HT含量显著性减少；相关调肝方药亦均可作用于上述靶点，有效纠正病变。这是愤怒、郁怒病机的共性所在之一。中缝背核GABABR1表达异常增多的程度不同可介导GABA调节中缝背核5-HT能神经元产生5-HT减少的程度也不同，这是愤怒与郁怒的一个不同点。由此说明，中缝背核GABABR1介导GABA调节5-HT生成是肝疏泄太过与不及的重要病机，也是“肝主疏泄调畅情志”微观机制之一。项目根据所用调肝方药的组分，推断白芍和香附的有效成分是发挥疗效作用的关键，为今后进一步探索提供了线索。