中文摘要：

目的探讨雌激素（E2）对雌激素受体-β（ER-β）与转录因子叉头蛋白3（FoxO3）的表达及其对卵巢颗粒细胞增殖与凋亡的影响。方法利用添加1μmol/L的E2或100 nmol/L ICI182.780的TCM199培养基体外培养卵巢颗粒细胞;采用WST比色法检测不同培养条件下卵巢颗粒细胞的增殖;RT-PCR和Real-time PCR检测ER-β与FoxO3 mRNA的表达,采用细胞免疫荧光和Western blotting检测ER-β与FoxO3的表达。结果 E2能促进卵巢颗粒细胞增殖,而ICI182.780抑制卵巢颗粒细胞增殖;RT-PCR扩增结果与理论值符合,Real-time PCR结果显示,ICI182.780与E2比较,ER-βmRNA表达下调,FoxO3 mRNA表达上调（P〈0.05）;Western blotting法结果显示,ER-β和FoxO3蛋白表达差异有统计学意义（P〈0.05）,灰度值分析显示添加E2中ER-β蛋白表达升高,FoxO3蛋白下降（P〈0.05）,添加ICI182.780中的ER-β表达下调,FoxO3表达上调,与E2比较差异有统计学意义（P〈0.05）。结论 E2可提高ER-β的表达,降低FoxO3的表达,影响卵巢颗粒细胞的增殖与凋亡。

英文摘要：

A Objective To explore the effects of estradiol （E2 ） on estrogen receptor-[3 （ER-[3） and forkhead transcription factor 3 （ FoxO3 ） expression in the proliferation and apoptosis of human granulosa cells during ovarian stimulation. Methods Ovarian granulosa cells in healthy ovarian follicles were cultured in TCM199 medium supplemented with 1 μmol/L E2, and 100 nmol/L ICI182. 780. The proliferation of ovarian granulosa cells was detected with WST. The ER-β and FoxO3 mRNA expressions in granulosa cells were determined with RT-PCR and real-time PCR. The ER-β and FoxO3 protein expressions were assessed with immunohistochemistry and Western blotting. Results E2 significantly improved while ICI182.780 inhibited the proliferation of ovarian granulosa cells. Real-time PCR results indicated that the ER-β mRNA expression was downregulated, while FoxO3 mRNA expression was upregulated （ P 〈 0.05）. Western blotting results showed that changes of ER-β and FoxO3 protein expressions were statistically significant （P 〈 0.05 ）. Grey level analysis showed that ER-β protein expression increased and FoxO3 protein expression was reduced with addition of E2 （P 〈 0.05 ）, while ER-β protein expression decreased and FoxO3 protein expression increased with addition of ICI182. 780 （ P 〈 0.05 ）. Conclusion E2 can affect the proliferation and apoptosis of ovarian granulosa cells by increasing ER-β expression while decreasing FoxO3 expression.