以肝脾为核心干预高催乳激素血症模型大鼠中枢多巴胺调控机制的研究

Hyperprolactinemia is the complicated case of reproductive axis dysfunction in gynecology. Currently ,it is not very clear impacting route and roling mechanism of central nervous system. Applicant deeply sduty this disease to regulatory mechanism of the follicular development, and research the dopamine channel initially ,so it provide the hypothesis that core to Ganpi treated intervene in the target to dopamine regulation mechanism of HPRL,which are base on viscera and meridian of theory and significance in clinical application with Shuganjianpi method .It adopt to modern technologies and new methods ,such as Western blot, RT-PCR，High performance liquid chromatography and so on.Through various respects to Morphology, Protein expression, Gene research to affect at different levels of dopamine rugulation (cyclase pathway of adenylate /Calcium signal transduction pathway /mitogeactivated protein kinase signal transduction pathway ),which verify the hypothesis in order to explore mechanism of dopamine regulation of HPRL,to research significant（useful） pathway of this disease to treat the liver and spleen as the core. Thinking of this study have not been reported in all over the world.To carry out the project will provide a theoretical basis so as to treat this disease of the liver and spleen as the core. To some extent,it enrich pathogenesis of HPRL.

高催乳激素血症是妇科生殖轴功能失调的疑难疾病，目前对中枢作用机制尚未完全清楚。项目申请人以肝脾藏象及经络理论为指导，在以疏肝健脾法临床运用疗效满意的基础上，深入进行了对本病卵泡发育影响的科研实践，并初步探索了多巴胺作用途径，进而提出了"以肝脾为核心干预高催乳激素血症模型动物中枢多巴胺调控机制"的假说，拟应用RT-PCR、Western blot、高效液相色谱法、免疫组化等现代新技术与新方法，采取分组比较，从形态学、基因水平、蛋白表达等不同层次研究多巴胺的调控机制（多巴胺腺苷酸环化酶通路、多巴胺钙离子信号转导通路、多巴胺MAPK信号转导通路）来验证假说，从而探讨高催乳激素血症影响中枢多巴胺调控的机制，探究以肝脾为核心治疗本病的作用途径。本研究思路国内外未见报道，项目实施将为以肝脾为核心论治本病提供理论依据，一定程度上丰富了高催乳激素血症的病机研究。

通过垂体移植造模方法最佳时间的探索，确定垂体移植后18天为制作HPRL模型的最佳时间，选择SCH23390 0.1mg/kg为最优多巴胺D1受体拮抗剂、氟哌啶醇1mg/kg为最优多巴胺D2受体拮抗剂；. 以肝脾为核心的治疗方法可以通过降低下丘脑DA的水平调节泌乳素的水平。通过检测 PRL激素的检测，来验证各治疗组的治疗效果。. 以肝脾为核心的治疗方法可以降低D1R、GS、升高D2R、Gi 、降低AC、CAMP、PKA、DARPP-32水平，推测其可能通过激动D2受体通路，抑制D1受体通路调节腺昔酸环化酶(AC)通路。. 以肝脾为核心的治疗方法可以上调PKC、IP3，上调Ca2＋浓度和CAMK水平，通过激活多巴胺D2受体钙离子通道调节泌乳素水平。. 以肝脾为核心的治疗方法可以下调LPS， PAK、 P38MAPK的水平，通过抑制D1受体P38通路调节泌乳素水平. 以肝脾为核心的治疗方法可以通过升高Ras、Raf、MEK1/2、ERK1/2的表达水平。通过激活D2受体ERK通路调节泌乳素水平. 实验验证了以肝脾为核心，运用逍遥散加减，可通过激动D2受体通路，抑制D1受体通路，从而达到调节HPRL的目的，丰富了中医药治疗HPRL的机制研究。