补肾助孕方对黄体功能不全模型大鼠垂体PKC/MAPK/ERK信号通路及黄体功能的影响  

Effect of Bushen Zhuyun Formula(补肾助孕方)on PKC/MAPK/ERK Signaling Pathway and Luteal Function in Luteal Phase Defect Model Rats

作　　者：曹珍珍[1] 周惠芳[1] CAO Zhenzhen;ZHOU Huifang(Nanjing University of Chinese medicine,Nanjing,210046)

机构地区：[1]南京中医药大学,南京210046

出　　处：《中医杂志》2021年第9期808-813,共6页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81774354,81473713);江苏省中医药局科技项目(ZD201702)。

摘　　要：目的探讨补肾助孕方治疗黄体功能不全(LPD)的可能作用机制。方法 40只雌性育龄SD大鼠随机分为正常组、模型组、补肾助孕方组、H-7组、H-7+补肾助孕方组,每组8只。除正常组外,其余各组大鼠均给予米非司酮混悬液建立LPD模型。补肾助孕方组(2.4g/100g灌胃)、H-7组(0.005mg/只腹腔注射)、H-7+补肾助孕方组(给药同补肾助孕方组和H-7组)均于第4个性周期(D13)开始,于D13、D17的16:00,及D14、D18的8:00给药,模型组、正常组均于以上给药时间点灌胃(1.2ml/100g)和腹腔注射(20μl/只)生理盐水。末次给药6h后,用ELISA法检测血清促黄体生成素(LH)、促卵泡生成素(FSH)含量,并计算FSH/LH值;HE法观察卵巢组织病理及卵泡、黄体数;Western blot法检测垂体组织蛋白激酶C(PKC)、磷酸化p38丝裂原活化蛋白酶(p-p38MAPK)、磷酸化细胞外信号调节激酶(p-ERK1/2)蛋白水平。结果与正常组比较,模型组和H-7+补肾助孕方组大鼠卵巢组织闭锁卵泡增多,黄体细胞发育不良,血清LH含量升高(P<0.05),卵巢组织卵泡数及黄体数,血清FSH含量及FSH/LH值,垂体组织PKC、p-p38MAPK、p-ERK1/2蛋白水平均降低(P<0.05);与模型组和H-7+补肾助孕方组比较,补肾助孕方组大鼠卵巢内闭锁卵泡减少,黄体细胞发育有所改善,血清LH含量降低(P<0.05),卵巢组织卵泡数及黄体数,血清FSH含量及FSH/LH值,垂体组织PKC、p-p38MAPK、p-ERK1/2蛋白水平升高(P<0.05)。结论补肾助孕方可激活垂体组织PKC/MAPK/ERK通路,促进血清FSH分泌,从而改善黄体功能。Objective To explore the possible mechanism of Bushen Zhuyun Formula(BZF,补肾助孕方) in treating luteal phase defect(LPD). Methods A total of 40 female SD rats of childbearing age were randomly divided into normal group, model group, BZF group, H-7 group and H-7 plus BZF group, with eight rats in each group. Except for the normal group, rats were given mifepristone suspension to establish the LPD model. Starting from the fourth sexual cycle(Day 13), BZF at a dose of 2.4 g/100 g was given by gavage in BZF group, while 0.005 mg H-7 was intraperitonealy injected into each rat in H-7 group at 16:00 on Day 13 and Day 17 and at 08:00 on Day 14 and Day 18;at the same time points, the rats in the H-7 plus BZF group were administered with same dose of drugs as the H-7 group and BZF group, and the rats in the model group and normal group were gavaged with 1.2 ml/100 g normal saline and intraperitonealy injected with 20μl normal saline per rat. At six hours after the last administration, the contents of luteinizing hormone(LH) and follicle stimulating hormone(FSH) in serum were detected by enzyme-linked immunosorbent assay(ELISA), and FSH/LH was calculated;hematoxylin eosin(HE) staining was used to observe the pathological changes of ovarian tissue and the numbers of follicles and corpus luteum;the protein levels of pituitary tissue protein kinase C(PKC), phosphorylated p38 mitogen activated protein kinases(p-p38 MAPK) and phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2) were detected by Western blotting. Results Compared to those in the normal group, the atresia follicle in the ovarian tissue increased in the model group and H-7 plus BZF group;there was dysplasia of corpus luteum;the content of serum LH increased(P<0.05);the numbers of ovarian follicles and corpus luteum, the content of serum FSH and FSH/LH ratio, and the protein levels of PKC, p-p38 MAPK and p-ERK1/2 in pituitary significantly increased(P<0.05). Compared to those in the model group and H-7 plus BZF group, the atresia follicle in th

关 键 词：黄体功能不全 补肾助孕方 蛋白激酶C 丝裂原激活蛋白激酶 胞外信号调节激酶 

分 类 号：R285.5[医药卫生—中药学]