电针通过Hippo/YAP通路调控卵巢早衰小鼠卵巢功能  

Electroacupuncture regulates ovarian function in mice with premature ovarian failure via Hippo/YAP pathway

作　　者：凌乐乐 王聪[2] 胡寅杰 李冰融 刘鹏[2] 张开勇[2] 李斯雯 刘特[3] 张必萌[2] LING Lele;WANG Cong;HU Yinjie;LI Bingrong;LIU Peng;ZHANG Kaiyong;LI Siwen;LIU Te;ZHANG Bimeng(Shanghai Jiao Tong University School of Medicine,Shanghai 200001,China;Shanghai General Hospital,Shanghai Jiao Tong University,Shanghai 200080,China;Shanghai Geriatrics Institute of Chinese Medicine,Shanghai 200030,China)

机构地区：[1]上海交通大学医学院,上海200001 [2]上海交通大学医学院附属第一人民医院,上海200080 [3]上海市中医老年医学研究所,上海200030

出　　处：《上海针灸杂志》2025年第2期229-238,共10页Shanghai Journal of Acupuncture and Moxibustion

基　　金：上海市进一步加快中医药事业发展三年行动计划2021年-2023年[ZY(2021-2023)-0208]。

摘　　要：目的观察电针改善高脂高糖诱导的卵巢早衰(premature ovarian failure,POF)小鼠卵巢功能与调控Hippo/YAP通路的分子机制。方法将45只C57BL/6雌性小鼠随机分为对照组(wild type,WT)、卵巢早衰组(POF)和电针治疗组(electroacupuncture,EA),每组15只。利用高脂高糖造模成功后,EA组选取关元穴及双侧足三里和三阴交穴进行电针干预。用苏木精-伊红(hematoxylin-eosin,HE)染色观察小鼠卵巢病理形态,用马松染色(Masson)观察小鼠卵巢纤维化,用菲律宾染色(filipin)检测小鼠卵巢总胆固醇;采用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测小鼠血清卵泡刺激素(follicle-stimulating hormone,FSH)、抗苗勒氏管激素(anti-Müllerian hormone,AMH)、雌二醇激素(estradiol,E2)、甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、高密度脂蛋白(high-density lipoprotein,HDL)和低密度脂蛋白(low-density lipoprotein,LDL)的水平;使用免疫荧光染色标记小鼠卵巢颗粒细胞(granulosa cells,GCs)卵泡刺激素受体(follicle-stimulating hormone receptor,FSHR)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和增殖标志物Ki67的水平;用逆转录定量实时荧光定量聚合酶链式反应(reverse transcription quantitative real-time polymerase chain reaction,RT-qPCR)检测Hippo/YAP通路基因YAP、TEAD1、TAZ、MST1、增殖相关基因PCNA、凋亡相关基因BAX、BCL2的mRNA表达;用Western blot法检测Hippo/YAP通路蛋白p-YAP、YAP、TEAD1、TAZ、MST1及凋亡相关蛋白BAX、BCL2的表达。结果与POF组比较,EA组干预后卵巢卵泡闭锁减少,成熟卵泡增多,卵巢纤维化程度减少,卵巢总胆固醇降低;EA组干预后性激素水平与脂代谢水平显著改善,FSH激素水平显著降低(P<0.05),AMH和E2激素水平显著升高(P<0.05),TG和TC显著下降(P<0.05),HDL显著上升(P<0.05),LDL显著下降(P<0.05);EA组干预后FSHR、PCNA、Ki67表达均上调;EA组干预后激活了Hippo/YAP通路,促进了GObjective To observe the molecular mechanism of electroacupuncture(EA)in improving ovarian function and regulating the Hippo/YAP pathway in mice with premature ovarian failure(POF)induced by high fat and high sugar.Method Forty-five female C57BL/6 mice were randomized into a control group(wild type,WT),a POF group,and an EA group,with 15 mice in each group.After the successful modeling with a high-fat and high-sugar diet,the EA group received EA at Guanyuan(CV4)and bilateral Zusanli(ST36)and Sanyinjiao(SP6)points.hematoxylin-eosin(HE)staining was adopted to observe ovarian pathological changes,Masson staining was used to observe ovarian fibrosis,and filipin staining was employed to detect ovarian total cholesterol in the mice.The enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of serum follicle-stimulating hormone(FSH),anti-Müllerian hormone(AMH),estradiol(E2),triglyceride(TG),total cholesterol(TC),high-density lipoprotein(HDL),and low-density lipoprotein(LDL).Immunofluorescence staining was used to label the expression of follicle-stimulating hormone receptor(FSHR),proliferating cell nuclear antigen(PCNA),and proliferation marker ki67 in the mice’s ovarian granulosa cells(GCs).The reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR)was adopted to detect the mRNA expression of Hippo/YAP pathway genes YAP,TEAD1,TAZ,and MST1,proliferation-related gene PCNA,and apoptosis-related genes BAX and BCL2.Western blot was used to measure the expression of Hippo/YAP pathway proteins p-YAP,YAP,TEAD1,TAZ,and MST1 and apoptosis-related proteins BAX and BCL2.Result Compared to the POF group,the EA group had less follicular atresia,more mature follicles,reduced ovarian fibrosis,and a lower ovarian TC level after the intervention.After the intervention,the EA group showed improvement in sex hormone and lipid metabolism levels:the FSH level dropped significantly(P<0.05),and the AMH and E2 levels increased significantly(P<0.05);the levels of TG and TC decreased markedly(P<0.05),

关 键 词：电针 卵巢早衰 Hippo/YAP通路 增殖 凋亡 小鼠 

分 类 号：R2-03[医药卫生—中医学]