雌激素对卵巢颗粒细胞雌激素受体-β和转录因子叉头蛋白3表达的影响  

Effect of estrogen on expression of estrogen receptor-β and forkhead transcription factor 3 in human granulosa cells

作　　者：马会明[1,2] 张永芳[1,3] 王蒙蒙[1] 李昕[1] 王永峰[1,3] 田洪成[1] 胡蓉[1,3] 王燕蓉[1,2] 裴秀英[1] 徐仙[1,3] 

机构地区：[1]宁夏医科大学生育力保持教育部重点实验室&宁夏回族自治区生殖与遗传重点实验室,宁夏银川750004 [2]宁夏医科大学人体解剖与组织胚胎学系组织胚胎学教研室,宁夏银川750004 [3]宁夏医科大学总医院生殖医学中心,宁夏银川750004

出　　处：《山东大学学报（医学版）》2016年第5期50-55,共6页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(31200882)

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。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.

关 键 词：雌激素 雌激素受体-Β 转录因子叉头蛋白3 卵巢颗粒细胞 

分 类 号：R711.6[医药卫生—妇产科学]