针刺对初老大鼠HPO轴及FSH/cAMP通路的影响  

Effects of acupuncture on the hypothalamic-pituitary-ovarian axis and FSH/cAMP signaling pathway in aged rats

作　　者：朱瑶瑶 尹雅倩 许焕芳[1] 杨莉[1] 黎蔚欣 苏晨晨 张嵘[3,4] 房繄恭[1] ZHU Yaoyao;YIN Yaqian;XU Huanfang;YANG Li;LI Weixin;SU Chenchen;ZHANG Rong;FANG Yigong(Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700,China;Affiliated Hospital of Hubei Provincial Government/Hubei Rehabilitation Hospital;Neuroscience Research Institute,Peking University;School of Basic Medical Sciences,Peking University)

机构地区：[1]中国中医科学院针灸研究所,北京100700 [2]湖北省直属机关医院/湖北省康复医院 [3]北京大学神经科学研究所 [4]北京大学基础医学院

出　　处：《中国针灸》2025年第2期200-208,共9页Chinese Acupuncture & Moxibustion

基　　金：国家重点研发计划项目:2022YFC3500504;国家自然科学基金资助项目:81674087。

摘　　要：目的:基于下丘脑-垂体-卵巢(HPO)轴整体调控及卵巢局部卵泡刺激素(FSH)/环磷酸腺苷(cAMP)信号通路2个角度,探讨针刺改善初老大鼠卵巢功能衰退的作用机制。方法:选取6只3月龄雌性SPF级SD大鼠作为空白组;另选取12只9月龄雌性SD大鼠随机分为模型组和针刺组,每组6只。针刺组大鼠于“百会”“关元”和双侧“次髎”行针刺干预,每次20 min,隔日1次,共针刺10次。每日采用阴道脱落细胞涂片观察大鼠动情周期;采用HE染色法观察大鼠卵巢组织形态,并对各级卵泡计数;ELISA法检测大鼠血清FSH、黄体生成素(LH)、雌二醇(E_(2))、抗缪勒管激素(AMH),下丘脑促性腺激素释放激素(GnRH),垂体FSH、LH及卵巢cAMP含量;免疫组化法及Westernblot法检测大鼠卵巢cAMP蛋白激酶催化亚基、促卵泡刺激素受体(FSHR)、P450蛋白表达;实时荧光定量PCR法检测卵巢FSHR、P450 mRNA表达。结果:与空白组比较,模型组大鼠动情周期紊乱率升高(P<0.01),卵泡颗粒细胞层减少、边界模糊、排列紊乱,各级发育卵泡减少、闭锁卵泡增多(P<0.01);血清FSH、LH含量升高(P<0.01),E_(2)、AMH含量降低(P<0.01);下丘脑Gn RH及垂体FSH、LH含量升高(P<0.01),卵巢cAMP含量降低(P<0.01);卵巢P450、cAMP蛋白激酶催化亚基、FSHR阳性表达及蛋白表达降低(P<0.01);卵巢FSHR、P450m RNA表达降低(P<0.01)。与模型组比较,针刺组大鼠动情周期紊乱率降低(P<0.01),颗粒细胞边缘清晰,原始卵泡和次级卵泡增多、闭锁卵泡减少(P<0.01);血清FSH、LH含量降低(P<0.01,P<0.05),E_(2)、AMH含量升高(P<0.05,P<0.01);下丘脑GnRH及垂体FSH、LH含量降低(P<0.01,P<0.05),卵巢cAMP含量升高(P<0.01);卵巢P450、cAMP蛋白激酶催化亚基、FSHR阳性表达及蛋白表达升高(P<0.01),卵巢FSHR、P450 mRNA表达升高(P<0.01)。结论:针刺可延缓初老大鼠卵巢功能衰退,其机制可能与调控HPO轴及FSH/c AMP信号通路有关。Objective To explore the mechanism of acupuncture on improving ovarian hypofunction in aged rats from two perspectives:the overall regulation of the hypothalamic-pituitary-ovarian(HPO)axis and the local ovarian follicle stimulating hormone(FSH)/cyclic adenosine monophosphate(cAMP)signaling pathway.Methods Six 3-month-old female SPF-grade Sprague-Dawley(SD)rats were selected as the blank group.Another twelve 9-month-old female SD rats were randomly divided into a model group and an acupuncture group,with six rats in each.The acupuncture group received acupuncture at"Baihui"(GV20),"Guanyuan"(CV4),and bilateral"Ciliao"(BL32)for 20 min per session,once every other day,for a total of 10 sessions.Vaginal smear tests were performed daily to observe the estrous cycle of the rats.Ovarian morphology was observed using HE staining,and follicles at various stages were counted.ELISA was used to detect levels of serum FSH,luteinizing hormone(LH),estradiol(E_(2)),anti-müllerian hormone(AMH),hypothalamic gonadotropin-releasing hormone(GnRH),pituitary FSH and LH,and ovarian cAMP.Immunohistochemistry and Western blot were used to detect the protein expression of ovarian cAMP protein kinase catalytic subunit,FSH receptor(FSHR),and P450.Real-time quantitative PCR was used to measure mRNA expression levels of FSHR and P450 in ovarian tissue.Results Compared with the blank group,the model group showed an increased rate of estrous cycle disorder(P<0.01),reduced granulosa cell layers with blurred boundaries and disordered arrangement,decreased numbers of developing follicles at all stages,and increased numbers of atretic follicles(P<0.01);the serum levels of FSH and LH were increased(P<0.01),while E_(2) and AMH levels were decreased(P<0.01);the hypothalamic GnRH and pituitary FSH and LH levels were elevated(P<0.01),and ovarian cAMP level was decreased(P<0.01);the positive expression and protein expression of ovarian P450,cAMP protein kinase catalytic subunit,and FSHR were reduced(P<0.01),and ovarian FSHR and P450 mRNA expression was de

关 键 词：卵巢功能衰退 针刺 卵泡刺激素(FSH)/环磷酸腺苷(cAMP)信号通路 下丘脑-垂体-卵巢(HPO)轴 

分 类 号：R245[医药卫生—针灸推拿学]