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

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(E₂)、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E₂、AMH水平上升，FSH水平下降(P<0.05或P&l...

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

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(E₂),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 E₂ and AMH levels were significantly increased(P<0.05 or P<0.01).The E₂ 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 E₂ 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 ERα expression in the Bushen Huoxue decoction group was more significant than that in the Bujiale group(P<0.05).Conclusion:POF model was successfully established by pZP3.The therapeutic mechanism of Bushen Huoxue decoction may be that it can enhance the biological effect of estrogen and mediate the process of disease recovery by increasing the expression of estrogen receptor in the ovary.