温补肾阳法调控AC-cAMP-PKA信号通路治疗抑郁症的作用机制研究

Depression is an important refractory disease of 21st century.After signal transmission of brain nerve cell receptors in the hippocampus is the hot topic of research of pathogenesis of depression and the effective mechanism of anti-depression drugs.Lots of research have found that the AC-cAMP-PKA signaling pathway of hippocampus has important role among the pathogenesis and therapeutic mechanism of depression. TCM can play a role in anti-depression via regulating this signaling pathway. Our pilot study has found that the pathogenesis of depression is complicated and varied,but kidney yang deficiency is one of the important pathogenesis.It can provide evidence for the TCM theory, clinic and experimental research. The basic formula(rou gui, ba ji tian) which based on kidney yang warming and tonfying method can improve the behavior performance of the animal model and regulate the disorder of 5-HT、DA、NE and CRH、ACTH、CORT. Kidney yang warming and tonfying method is getting more and more popular as one of the effective therapeutic methods, it should be researched intensively. But there is no research on the effective mechanism of kidney yang warming and tonfying method form the perspective of AC-cAMP-PKA signaling pathway. This program will adopt the CUMS method to establish the depression rat model, using the method of elisa、RIA、Western blotting、RT-PCR,ect，testing expression of AC、cAMP、PKA，CREB 、BDNFmRNA to observe the affection of formula(rou gui, ba ji tian) which based on kidney yang warming and tonfying method on the AC-cAMP-PKA signaling pathway of hippocampus and explore the effective mechanism of kidney yang warming and tonfying method, which provide new thinking for TCM treatment and prevention of depression.

抑郁症是21世纪重大难治病，海马组织受体后信号传导是目前研究抑郁症发病机制和抗抑郁药作用机制的热点，其中AC-cAMP-PKA信号通路是关键路径。近年温补肾阳法治疗抑郁症显示出重要的临床与学术价值。前期研究发现：温补肾阳法能有效改善抑郁症模型动物行为学表现及5-HT、DA、NE和CRH、ACTH、CORT紊乱，且还能调节AC-cAMP-PKA信号通路上的重要中间环节cAMP水平，提示温补肾阳法可能通过调控AC-cAMP-PKA信号通路系统发挥抗抑郁作用，但目前尚缺乏相关研究。本项目采用国际公认的CUMS法建立大鼠抑郁症模型，运用elisa、RIA、Western blotting、RT-PCR等技术，检测AC、cAMP、PKA蛋白表达，CREB 、BDNFmRNA表达等，揭示以温补肾阳立法的复方（肉桂、巴戟天）调控AC-cAMP-PKA信号通路治疗抑郁症的作用机制，探明该方药物作用环节。

抑郁症是21世纪重大难治病，海马组织受体后信号传导是目前研究抑郁症发病机制和抗抑郁药作用机制的热点，其中AC-cAMP-PKA信号通路是关键路径。近年温补肾阳法治疗抑郁症显示出重要的临床与学术价值。本项目采用国际公认的CUMS法建立大鼠抑郁症模型，用旷野试验进行评估造模。酶联免疫吸附法（Elisa）测定大鼠海马AC、cAMP、5-HT、NE、DA、5-HIAA含量，RT-PCR法检测大鼠海马CREB mRNA、BDNF mRNA表达，蛋白质印迹法（Western blotting）检测海马组织PKAc蛋白水平，并增加了对大鼠大脑组织的光镜下观察，确定大鼠抑郁模型的病理变化及治疗效果，揭示以温补肾阳立法的复方（肉桂、巴戟天）调控AC-cAMP-PKA信号通路治疗抑郁症的作用机制。实验结果显示，温补肾阳方可以上调大鼠海马AC-cAMP-PKA信号通路，明显提高大鼠海马5-HT、NE、DA含量，上调大鼠海马CREB mRNA和BDNF mRNA表达，改善抑郁大鼠脑组织病理变化，提示临床可采用温补肾阳法治疗抑郁症发挥抗抑郁作用。温补肾阳法治疗抑郁症可通过调控AC-cAMP-PKA信号通路而实现，为临床使用温补肾阳法治疗抑郁症奠定了一定的学术思想，并提供了实验基础和理论依据，发表学术论文6篇，参编相关论著1部，参加学术交流会5次，申请专利1项（已获初审通过），继续申报中标厅局级课题1项，培养青年骨干6名，硕士研究生5名。