论文部分内容阅读
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 observe the effect of low dose mifepristone on cultured human ovarian granulosa cells in vitro and to explore its molecular mechanism of inhibiting or delaying ovulation. Methods The effects of mifepristone on the morphology and ultrastructure of cultured human ovarian granulosa cells were observed by inverted microscope and electron microscope. Immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR) ) Were used to detect the effects of different concentrations of mifepristone on the expression of Bcl-2 and Bax protein and mRNA in cultured granulosa cells. Results Mifepristone induced the apoptosis of human ovarian granulosa cells. After treatment with different concentrations of mifepristone (2.5μmol / L, 5μmol / L, 10μmol / L) for 24 hours, the expression of Bcl- (HSCORE) were 2.15 ± 0.16, 1.88 ± 0.13 and 1.64 ± 0.16, respectively, which were significantly different from those of the control group (P <0.001), while the expression of Bax was opposite HSCORE were 1.85 ± 0.10, 2.00 ± 0.18 and 2.29 ± 0.20, respectively, which were significantly different from those of control group (P <0.001), while the mRNA expression of bcl-2 and bax was not significantly different from that of the control group Change (P> 0.05). Conclusion Mifepristone can induce the apoptosis of human ovarian granulosa cells. The expression of Bcl-2 and Bax may be involved in the apoptosis of granulosa cells induced by mifepristone.