低剂量米非司酮对人卵巢颗粒细胞的影响  

EFFECTS OF LOW-DOSE MIFEPRISTONE ON HUMAN GRANULOSA CELLS

作　　者：郭庆云[1] 张武文[1] 庄亚玲[1] 黄丽丽[1] 

机构地区：[1]浙江大学医学院附属妇产科医院,杭州310006

出　　处：《解剖学报》2008年第6期872-876,共5页Acta Anatomica Sinica

基　　金：浙江省科技厅计划资助项目(2003C33013)

摘　　要：目的观察低剂量米非司酮对体外培养的人卵巢颗粒细胞的影响,探讨其抑制或延迟排卵的分子机制。方法应用细胞培养,通过倒置显微镜、电子显微镜观察米非司酮对体外培养的人卵巢颗粒细胞形态及超微结构的影响;用免疫细胞化学和反转录-聚合酶链式反应(RT-PCR)分别检测不同浓度米非司酮处理24h后对颗粒细胞Bcl-2和Bax蛋白及其mRNA表达水平的影响。结果米非司酮可以诱导人卵巢颗粒细胞发生凋亡;不同浓度米非司酮(2.5μmol/L、5μmol/L、10μmol/L)处理24h后,颗粒细胞Bcl-2蛋白表达水平进行组织化学评分(HSCORE),分别为2.15±0.16、1.88±0.13和1.64±0.16,与对照组的2.51±0.16比较,差异有显著性(P<0.001);Bax蛋白表达则出现相反的变化,3组的HSCORE分别为1.85±0.10、2.00±0.18和2.29±0.20,与对照组的1.50±0.16比较,差异有显著性(P<0.001);而bcl-2和bax基因mRNA表达与对照组相比无明显改变(P>0.05)。结论米非司酮能诱导人卵巢颗粒细胞凋亡;Bcl-2和Bax蛋白表达可能参与米非司酮诱导的颗粒细胞凋亡。Objective To explore the effects of low-dose mifepristone on human granulosa cells and the molecular mechanism of its inhibiting or delaying ovulation. Methods Phase contrast microscopy and electron microscopy were conducted to observe the morphological and biochemical change of granulosa cells exposed to various doses of mifepristone for 24 hours （the final concentrations of mifepristone were respectively 2.5μmol/L, 5μmol/L and 10μmol/L, Non-mifepristone as the control group）. Immunocytochemistry was performed to detect the protein expressions of Bcl-2 and Bax in granulosa cells exposed to various doses of mifepristone for 24 hours. The mRNA levels of Bcl-2 and Bax in granulosa cells were detected by RT-PCR. Results Granulosa cells treated with various doses of mifepristone presented typical morphological characteristics of apoptosis such as condensation of chromatin and cytosolic, and shrink of cells. The expression of Bcl-2 protein might be down-regulated, while the expression of Bax might be up-regulated by mifepristone. The histological score （HSCORE） of Bcl-2 protein in granulosa cells, which were incubated with various doses of mifepristone （2.5μmol/L, 5μmol/L, 10μmol/L） for 24 hours, were respectively 2.15 ± 0.16,1.88 ±0.13 and 1.64 ±0.16 respectively, Compared with the HSCORE of the control group （2.51 ± 0.16）, the difference was significant （P 〈 0.001 ）. Compared with the control group （1.50 ± 0.16）, there was singificant difference in the expression of Bax protein （ 1.85 ± 0.10, 2.00 ＋ O. 18 and 2.29 ±0.20） （ P 〈 0. 001 ）. However, the mRNA levels of bcl-2 and bax after 24 hours incubation with various doses of mifepristone （ P 〉 O. 05 ）. Conclusion Mifepristone could induce human granulosa cells apoptosis. The Bcl-2 protein and Bax protein may be involved in the apoptosis of granulosa cells induced by mifepristone.

关 键 词：米非司酮 颗粒细胞 BCL-2 BAX 细胞培养 免疫组织化学 反转录-聚合酶链式反应 人 

分 类 号：R711[医药卫生—妇产科学]