补肾助孕方治疗LPD性不孕症的新机制：垂体GnRH受体后信号通路

项目名称： 补肾助孕方治疗LPD性不孕症的新机制：垂体GnRH受体后信号通路

项目编号： No.81473713

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 周惠芳

作者单位： 南京中医药大学

项目金额： 74万元

中文摘要： 补肾助孕方是基于名老中医夏桂成教授提出的心(脑)-肾-子宫轴理论指导下形成的经验方剂。临床研究已证实其为治疗黄体功能不全（LPD）性不孕症的有效方药，并能提高患者胚胎着床期P、LH等水平；实验结果提示其改善卵巢黄体功能及子宫内膜容受性。基于中西医对LPD性不孕为生殖轴功能失调认识上的一致性，我们认为补肾助孕方能够作用于垂体，提出通过GnRH受体后信号通路调控垂体GTH表达进而调节生殖轴功能的作用新机制假说。本项目以米非司酮构建LPD大鼠模型、西曲瑞克构建GnRH受体拮抗细胞模型，采用qPCR、Western Blot、ELISA、免疫荧光共聚焦扫描等细胞分子生物学技术，首次从GnRH受体后PKC-MAPK、Ca2+-CAM、cAMP-PKA信号通路，开创性研究补肾助孕方对垂体GTH表达分泌的调控及补肾法治疗LPD的助孕新机制，为中药治疗LPD不孕症的研究提供新思路、新方法。

中文关键词： 补肾助孕方；黄体功能不全；促性腺激素；GnRH受体；信号通路

英文摘要： The Prescription for Reinforcing Kidney and Promoting Pregnancy is an epirical formula formed under the theoretical guidance of the 'Heart(Brain)-Kidney-Uterus axis' proposed by Xia Guicheng, the renowned specialist of TCM. Previous clinical researches have confirmed its efficacy in the treatment of luteal phase defect (LPD) infertility, which can increase patients' progesterone (P) and luteal hormone (LH) levels during the embryo implantation stage while experimental results suggest that the prescription can improve the corpus luteum's function and endometrial receptivity. Based on the consistency of both Chinese and western medicine's understandings that the LPD infertility is due to the dysfunction of the reproductive axis, we consider that the effective ingredients of the formula can act on the pituitary directly. Therefore, we are proposing a hypothesis about a new mechanism that the prescription can regulate the expression of GTH through the signaling pathway via GnRH receptor to adjust the function of the reproductive axis. In this project, we will study the rat LPD model with Mifepristone and the pituitary cell GnRH receptor antagonistic model with Cetrorelix，and adopt cellular and molecular biology techniques such as qPCR、Western Blot、ELISA and Immunofluorescence laser scanning confocal microscope（IF/LSCM）. For the first time, on the basis of the PKC-MAPK、Ca2+-CAM and cAMP-PKA signaling pathway via GnRH receptor, this project will innovatively explore the new pregnancy-promoting mechanism of the Prescription for Reinforcing Kidney and Promoting Pregnancy in regulating the expression and secretion of GTH and the kidney-tonifying methods in curing LPD infertility. Thus, we will discover new thoughts and methods for the research of treating LPD infertility with Chinese medicine.

英文关键词： the Prescription for Reinforcing Kidney and Promoting Pregnancy;luteal phase defect（LPD）;gonadotropic hormone(GTH);GnRH receptor;signaling pathway