补肾促排方对大鼠卵巢颗粒细胞黄体生成素受体和卵泡刺激素受体的影响

Influence of Bushen Cupai formula on the expression of luteinizing hormone receptor and follicle stimulating hormone receptor in rat ovarian granulosa cells

目的通过体外实验阐述补肾促排方治疗多囊卵巢综合征的药理学作用机制。方法提取与鉴定大鼠卵巢颗粒细胞,提取补肾促排方水溶液。实验分为补肾促排方低、中、高剂量组和空白对照组,分别相应处理分离与鉴定的大鼠卵巢颗粒细胞24 h后提取mRNA,48 h后提取蛋白。通过RT-PCR和Wester-blot法分别检测各组黄体生成素受体(LHR)和卵泡刺激素受体(FSHR)mRNA和蛋白表达水平。结果补肾促排方水溶液能够显著上调大鼠卵巢颗粒细胞LHR和FSHR mRNA和蛋白表达水平(P均<0.05),且呈现显著的浓度梯度依赖效应。结论补肾促排方通过上调卵巢颗粒细胞LHR和FSHR的表达,促进卵泡的发育与排出,从而起到治疗多囊卵巢综合征的作用。

Objective It is to elucidate the underlying mechanism of pharmacology of Bushen Cupai formula in the treatment of polycystic ovary syndrome by experiment in vitro. Methods The rat ovarian granulosa cells were isolated and authenticated,Bushen Cupai formula was made into liquid fomula. Rat ovarian granulosa cells were treated with low,medium and high doses of Bushen Cupai formula and its empty control. mRNA was isolated after 24 h treatment and protein was extracted after 48 h treatment. RT-PCR and Western-blot were utilized to check the expression of luteinizing hormone receptor（LHR）and follicle stimulating hormone receptor（FSHR） at mRNA level and protein level separately. Results Bushen Cupai formula significantly increased the expression of LHR and FSHR both in mRNA level and protein level in a concentration-dependent manner（P〈 0. 05）. Conclusion Bushen Cupai formula combated polycystic ovary syndrome by up-regulating LHR and FSHR,which promoted the development and releasing of follicle as a consequence.