补肾助孕方对LPD模型大鼠垂体GnRH受体信号转导系统的影响  

Effect of Bushen Zhuyun Formula on Pituitary GnRH Receptor and Signal Transduction System in LPD Rats

作　　者：周伯如[1] 周惠芳[1] 刘贝[1] 朱宏坤 戴佳轩 徐建亚[1] 谢彤[1] 

机构地区：[1]南京中医药大学第一临床医学院,江苏南京210023

出　　处：《南京中医药大学学报》2017年第6期554-559,共6页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81473713);江苏省自然科学基金(BK20141463);江苏省中医药管理局重点项目(ZD201702)

摘　　要：目的通过观察补肾助孕方对大鼠垂体促性腺激素及促性腺激素释放激素(GnRH)受体信号系统的影响,探究其作用机制,寻找中医心-肾-子宫轴理论的现代科学依据。方法采用米非司酮对SD雌鼠进行造模,并进行药物干预。采用ELISA法检测大鼠血清样本促黄体生成激素(LH)和促卵泡成熟激素(FSH)水平,qPCR法检测各组大鼠垂体FSHβ、LHβ基因转录水平。对大鼠垂体GnRH受体信号转导系统关键分子以及下游重要转录因子分别采用qPCR与Western blot方法检测其基因转录与蛋白表达水平。结果补肾助孕方显著提高了由米非司酮诱导的血清低FSH水平与垂体GnRH受体低表达(P<0.05),补肾助孕方组高、低剂量组中PKC、p38MAPK、JNK、ERK、PKA等关键信号转导分子以及c-Jun、Elk-1、Egr-1、CREB等转录因子的mRNA及蛋白表达均显著高于模型组(P<0.05),但CAM及其下游的Nur77在各组间均无显著差异。结论补肾助孕方能够通过调节垂体GnRH受体信号转导系统调节垂体促性腺激素的分泌水平,证明该方具有调节生殖轴整体功能的作用,阐释了中医心-肾-子宫轴理论与现代医学下丘脑-垂体-卵巢轴在内容上的一致性。OBJECTIVE To explore the effect and mechanism of Bushen Zhuyun formula(BSZYF)on gonadotropins and gonadotropin-releasing hormone receptor(GnRHR)signaling transduction in mifepristone-induced LPD rats,and to find the scientific evidence of 'heart,kidney and uterus axis' theory of TCM.METHODS SD female rats were administered mifepristone suspension via oral gavage,followed by pharmaceutical intervention.Then,the serum follicle-stimulating hormone(FSH)and luteinizing hormone(LH)levels were detected by ELISA assay,the FSHβand LHβexpression levels in the pituitary were determined by qPCR,and the GnRHR expression levels were assessed by qPCR and Western blot analysis.Simultaneously,transcription factors,such as c-Jun,Elk-1,Egr-1,Nur77 and CREB,and transduction molecules in PKC-MAPK,Ca2+-CAM,and cAMP-PKA signaling pathways,such as PKC,p38 MAPK,ERK1/2,CAM,and PKA were assessed by qPCR and Western blot analysis.RESULTS After pharmaceutical intervention,BSZYF increased the serum FSH levels suppressed by mifepristone(P<0.05).Moreover,the expression of c-Jun,Elk-1,Egr-1,CREB,PKA,p38 MAPK,JNK,ERK and PKA were was significantly upregulated in the BSZYF group compared with that in the mifepristone group(P<0.05).However,there was no statistical difference in Nur77 and CAM expression among the blank,mifepristone and BSZYF groups(P>0.05).CONCLUSION BSZYF can regulate abnormal pituitary gonadotropin secretion and GnRHR signaling transduction systems induced by mifepristone,which confirms that the prescription can regulate the whole function of the reproductive axis,and explains the consistency of ' Heart,kidney and uterus axis' theory in TCM and the concept of the hypothalamus-pituitary-ovary axis in modern medicine

关 键 词：补肾助孕方 黄体功能不全 信号通路 GNRH受体 

分 类 号：R285.5[医药卫生—中药学]