二十五味鬼臼丸的植物雌激素样作用及其对雌激素受体的调控

Roles of Ershiwuwei Guijiu Pills in generating phytoestrogen-like effects and regulating estrogen receptors

目的探讨二十五味鬼臼丸对围绝经期综合征大鼠模型植物雌激素样作用及其对雌激素受体的调控。方法采用切除双侧卵巢的方法构建围绝经期综合征大鼠模型,将60只SD雌性大鼠随机分为假手术组、模型组、雌二醇组和二十五味鬼臼丸组,每组15只。围绝经期综合征模型建立成功后,假手术组和模型组给予等容量生理盐水,雌二醇组给予0.1 mg/kg戊酸雌二醇,二十五味鬼臼丸组给予0.36 g/kg二十五味鬼臼丸,隔日灌胃,连续8周。计算子宫指数(UI),ELISA检测血清雌二醇(E_(2))、黄体生成素(LH)和卵泡刺激素(FSH)水平,RT-qPCR检测子宫组织中雌激素受体α(ERα)、雌激素受体β(ERβ)mRNA表达,Western blot及免疫荧光染色法检测子宫ERα、ERβ蛋白表达。结果与假手术组比较,模型组子宫指数、血清E_(2)水平降低(P<0.01),LH、FSH水平升高(P<0.01),子宫ERα、ERβmRNA及蛋白表达降低(P<0.01);与模型组比较,二十五味鬼臼丸组子宫指数、血清E_(2)水平升高(P<0.01),LH、FSH水平降低(P<0.05),ERβmRNA及蛋白表达升高(P<0.05,P<0.01),子宫ERαmRNA及蛋白表达无明显变化(P>0.05)。结论二十五味鬼臼丸在子宫组织中选择性上调ERβ的表达,对ERα无明显影响,表明其具有选择性雌激素受体调节剂作用,而选择性雌激素受体调节作用可能是二十五味鬼臼丸有效改善围绝经期综合征的分子机制之一。

AIM To investigate the phytoestrogen-like effects of Ershiwuwei Guijiu Pills on perimenopausal syndrome rats and its role in the regulation of estrogen receptors.METHODS Bilateral ovaries excision was performed in constructing rat model of perimenopausal syndrome. A total of sixty SD female rats were randomly divided into sham operation group, model group, estradiol group and Ershiwuwei Guijiu Pills group, with fifteen rats in each group. After the successful establishment of perimenopausal syndrome rat models, 8-week oral gavage of same amount of normal saline in the sham operation group and the model group, 0.1 mg/kg estradiol valerate in the estradiol group, and 0.36 g/kg Ershiwuwei Guijiu Pills in Ershiwuwei Guijiu Pills group were carried out every other day. The rats were subjected to uterine index(UI) level calculation;measurement of serum levels of estradiol(E_(2)), luteinizing hormone(LH) and follicle-stimulating hormone(FSH) by ELISA;detection of mRNA expressions of estrogen receptor α(ERα) and estrogen receptor β(ERβ) in uterine tissues by RT-qPCR;and determination of expressions of ERα and ERβ protein in uterine tissue by Western blot and immunofluorescence staining.RESULTS Compared with the sham group, the model group shared decreased uterine index level(P<0.01) and serum E_(2)level(P<0.01), increased LH and FSH levels(P<0.01), and decreased protein expressions of ERα, ERβ mRNA and proteins(P<0.01). Compared with the model group, Ershiwuwei Guijiu Pills group demonstrated increased UI level(P<0.05), increased serum E_(2)level(P<0.01), decreased LH and FSH levels(P<0.05), and increased expression of ERβ mRNA and protein(P<0.05, P<0.01), but with no significant difference in the expression of ERα mRNA and protein(P>0.05).CONCLUSION Ershiwuwei Guijiu Pills’ selective effect in increasing ERβ expression instead of ERα expression in uterine tissue, indicates their role as estrogen receptor modulator contributing the effective improvement of perimenopausal syndrome.