补肾生精调和气血方调控Wnt/β-catenin信号通路对卵巢储备功能减退大鼠的影响  

Effects of Bushen Shengjing Tiaohe Qixue Prescription in Regulating Wnt/β-catenin Signaling Pathway on Diminished Ovarian Reserve Rats

作　　者：王珺 唐娜 叶朝阳[1,2] 黎玲 高同庆 黄佳梅 邓强 吴锋英 金志春 WANG Jun;TANG Na;YE Chaoyang;LI Ling;GAO Tongqing;HUANG Jiamei;DENG Qiang;WU Fengying;JIN Zhichun(Hubei University of Chinese Medicine,Wuhan 430065,China;Maternal and Child Health Hospital of Hubei Province,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430070,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]华中科技大学同济医学院附属湖北妇幼保健院,湖北武汉430070

出　　处：《中国中医药信息杂志》2025年第4期100-106,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：全国名老中医药专家传承工作室建设项目(2022年);湖北省卫生健康委中医药科研重点项目(ZY2021Z013)。

摘　　要：目的基于Wnt/β-catenin信号通路观察补肾生精调和气血方对卵巢储备功能减退(DOR)大鼠的影响。方法60只8周龄SD雌性大鼠随机分为对照组、模型组、戊酸雌二醇组和中药低、中、高剂量组,每组10只。除对照组外,其余组单次腹腔注射环磷酰胺75 mg/kg复制DOR大鼠模型。造模次日开始,给药组分别予相应药液灌胃,连续3周。观察大鼠一般状况,ELISA检测血清卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E_(2))、抗米勒管激素(AMH)、抑制素B(INHB)含量,HE染色观察卵巢组织形态,Western blot检测卵巢组织糖原合成酶激酶3β(GSK3β)、β-连环蛋白(β-catenin)、细胞周期蛋白D1(CyclinD1)、c-MYC蛋白表达,免疫组化染色检测卵巢组织β-catenin蛋白表达,RT-qPCR检测卵巢组织Wnt1、Wnt2、Wnt4、GSK3β、β-catenin、CyclinD1、c-MYC mRNA表达,透射电镜观察卵巢颗粒细胞超微结构。结果与对照组比较,模型组大鼠进食饮水量减少,体质量增长缓慢甚至降低,动情周期紊乱,血清FSH、LH含量升高(P<0.01),E_(2)、AMH、INHB含量降低(P<0.01);卵巢组织各级卵泡数量减少、闭锁卵泡增多,颗粒细胞排列紊乱;卵巢组织GSK3β蛋白表达升高(P<0.01),β-catenin、CyclinD1、c-MYC蛋白表达降低(P<0.05,P<0.01),β-catenin阳性表达降低(P<0.01),卵巢组织Wnt1、Wnt2、Wnt4、β-catenin、CyclinD1、c-MYC mRNA表达降低(P<0.01),GSK3βmRNA表达升高(P<0.01);卵巢颗粒细胞线粒体、粗面内质网等细胞器高度肿胀,部分溶解。与模型组比较,各给药组大鼠体质量不同程度增加,动情周期部分恢复,血清FSH、LH含量降低,E_(2)、AMH、INHB含量升高(P<0.01,P<0.05);卵巢组织各级卵泡数量不同程度增多、闭锁卵泡减少;卵巢组织GSK3β蛋白表达降低,β-catenin、CyclinD1、c-MYC蛋白表达升高,β-catenin阳性表达升高,卵巢组织Wnt1、Wnt2、Wnt4、β-catenin、CyclinD1、c-MYC mRNA表达升高,GSK3βmRNA表达降低,其中戊酸雌二醇组�Objective To investigate the effects of Bushen Shengjing Tiaohe Qixue Prescription on rats with diminished ovarian reserve(DOR)based on Wnt/β-catenin signaling pathway.Methods Totally 608-week-old SD female rats were randomly divided into control group,model group,estradiol valerate group and TCM low-,medium and high-dosage groups,with 10 rats in each group.Except for the control group,the rats in the other groups were given a single intraperitoneal injection of cyclophosphamide 75 mg/kg to replicate the DOR rat model.From the next day of modeling,the corresponding drugs was administered for medication groups by gavage for 3 weeks.The general conditions of the rats were observed,ELISA was used to detect the contents of serum follicle-stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E_(2)),anti-mullerian hormone(AMH)and inhibin B(INHB),HE staining was used to observe the morphologic changes in ovarian tissue,Western blot was used to detect GSK3β,β-catenin,CyclinD1 and c-MYC protein expressions in ovarian tissues,immunohistochemical staining was used to observeβ-catenin expression in ovarian tissue,RT-qPCR was used to detect Wnt1,Wnt2,Wnt4,GSK3β,β-catenin,CyclinD1 and c-MYC mRNA expression in ovarian tissue,transmission electron microscopy was used to observe ultrapathologic structure of ovarian granulosa cells.Results Compared with the control group,the rats in the model group had reduced intake of food and water,slow or even reduced body mass gain,and disturbed estrous cycle,the serum contents of FSH and LH increased(P<0.01),the contents of E_(2),AMH and INHB decreased(P<0.01);the number of follicles of all levels in ovarian tissue was reduced,and there was increased number of atretic follicles with disturbed arrangement of the granulosa cells;the expression of GSK3βprotein in ovarian tissue increased(P<0.01),the expression ofβ-catenin,CyclinD1 and c-MYC protein decreased(P<0.05,P<0.01),andβ-catenin positive expression decreased(P<0.01),the expression of Wnt1,Wnt2,Wnt4,β-catenin,CyclinD1 and c

关 键 词：补肾生精调和气血方 卵巢储备功能减退 WNT/Β-CATENIN信号通路 细胞周期蛋白D1 C-MYC 大鼠 

分 类 号：R285.5[医药卫生—中药学]