基于雌激素受体途径探讨补肾活血汤干预免疫性卵巢早衰小鼠模型机制研究  

Investigation of the mechanism of intervention in immune premature ovarian failure mouse model with Bushen Huoxue decoction based on estrogen receptor pathway

作　　者：陈玥 陈思[1] 张蕾[1] 钟伟萍[1] 佘婧瑶 卢燕[1] 王佩娟[1] 王静[1] CHEN Yue;CHEN Si;ZHANG Lei;ZHONG Weiping;SHE Jingyao;LU Yan;WANG Peijuan;WANG Jing(Department of Obstetrics and Gynecology,Affiliated Hospital of Integrated Traditional Chinese and Western Medicine,Nanjing 210028,China)

机构地区：[1]江苏省中西医结合医院妇产科,江苏南京210028 [2]南京中医药大学第三临床医学院,江苏南京210023

出　　处：《陕西中医》2023年第5期556-561,共6页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81903998,81904245,82074487);中国中西医结合妇产科专项研究基金普正制药专项课题(FCK-PZ-09);江苏省名老中医药专家王佩娟传承工作室建设项目(20SG2S001)。

摘　　要：目的:以透明带3多肽(pZP3)诱导的免疫性卵巢早衰(POF)小鼠为研究对象,观察其血清性激素水平及卵巢组织中雌激素受体(ER)表达情况,探讨免疫性卵巢早衰可能的发生机制及补肾活血汤的潜在治疗途径。方法:将40只雌性B6AF1小鼠随机分为空白组、模型组、补佳乐组及补肾活血汤组。除空白组外,各组小鼠第1天、第14天两次予pZP3联合弗氏佐剂皮下注射造模,空白组同期予0.9%氯化钠溶液皮下注射;建模成功后空白组、模型组、补佳乐组及补肾活血汤组分别予以等体积0.9%氯化钠溶液、戊酸雌二醇及补肾活血汤灌胃,每天2次。灌胃28 d后处死小鼠,采用酶联免疫吸附测定法(ELISA)检测血清卵泡刺激素(FSH)、雌二醇(E_(2))、抗缪勒氏管激素(AMH)水平;采用HE染色和免疫组化方法检测卵巢组织病理变化及雌激素受体α(ERα)、雌激素受体β(ERβ)的表达情况。结果:①与空白组相比,模型组小鼠呈现出典型的低雌激素高促性腺激素表型(P<0.01);经补佳乐及补肾活血汤灌胃治疗后,小鼠血清E_(2)、AMH水平上升,FSH水平下降(P<0.05或P<0.01),补佳乐组雌激素上升水平更加显著(P<0.05)。②模型组中,小鼠卵巢中卵泡数目减少,闭锁卵泡增多,ER表达与E_(2)水平下降呈现平行趋势(P<0.01);予补佳乐及补肾活血汤灌胃后,小鼠卵巢中各级卵泡恢复发育,雌激素受体表达明显上调(P<0.01),补肾活血汤组小鼠卵巢中ERα的表达上调较补佳乐组更加显著(P<0.05)。结论:pZP3造模可有效构建POF模型,补肾活血汤的治疗机制可能是通过提高卵巢雌激素受体表达,从而增强雌激素的生物学效应的发挥,介导疾病恢复过程。Objective:Immune premature ovarian failure(POF)mice induced by zona pellucida 3 polypeptide(pZP3)were recruited to investigate the serum levels of sex hormones and the expression of estrogen receptor(ER)in ovarian tissue,as well as to explore the possible mechanism of immune POF and the potential therapeutic pathway of Bushen Huoxue decoction.Methods:Forty female B6AF1 mice(6-8 weeks old)were randomly divided into control group,model group,Bujiale group,and Bushen Huoxue decoction group.Two injections of pZP3 combined with Freund’s adjuvant subcutaneously on days 1 and 14 in each group except the control,and 0.9%saline subcutaneously during the same period in the control group.After successful modeling,the control group,model group,Bujiale group,and Bushen Huoxue decoction group were treated with equal volumes of 0.9%saline,estradiol valerate,and Bushen Huoxue decoction by gavage once a day.Mice were sacrificed after 28 days of gavage,and serum levels of the follicular stimulating hormone(FSH),estradiol(E_(2)),and anti-Mullerian hormone(AMH)were assessed by ELISA.The ovarian pathology and the expression of estrogen receptor changes were observed by HE staining and immunohistochemistry.Results:①Compared with the control group,the model mince was characterized by low estrogen and high gonadotropin levels(P<0.01).After treatment with estradiol valerate and Bushen Huoxue decoction,the serum levels of FSH were reduced considerably,while the E_(2)and AMH levels were significantly increased(P<0.05 or P<0.01).The E_(2)levels in Bujiale group were more obviously(P<0.05).②In the model group,the number of follicles in the ovaries decreased,the number of atretic follicles increased,and the ER expression showed a parallel trend with the decrease in E_(2)level(P<0.01).The development of follicles at all levels was restored,and the expression of estrogen receptor in the ovary of mice was significantly up-regulated after gavage with Bujiale and Bushen Huoxue decoction(P<0.01).The up-regulation of ERαexpression in the Bush

关 键 词：免疫性卵巢早衰 透明带3多肽 雌激素受体 补肾活血汤 小鼠模型 免疫组化 

分 类 号：R711.75[医药卫生—妇产科学]