米非司酮对人子宫内膜细胞凋亡及促血管生成因子表达的影响

Effect of Mifepristone on Expression of Angiogenic Factors and Apoptosis of Human Endometrial Cells

目的:探讨米非司酮对人子宫内膜细胞凋亡以及促血管生成因子血管生成素1(Ang 1)、血管生成素2(Ang2)、血管内皮生长因子(VEGF)和碱性成纤维细胞因子(b FGF)表达水平的影响。方法:采用MTS法检测不同浓度米非司酮对子宫内膜细胞的毒性,采用Annexin V-FITC/PI染色流式细胞术和Western Blot检测细胞凋亡,并用实时荧光定量聚合酶链反应检测促血管生成因子的表达。结果:米非司酮浓度增加,细胞活力随之减小;米非司酮作用时间增长,细胞活力随之减小,24、48、72小时的IC 50分别为18、15、12μg/ml。与对照组相比,不同浓度的米非司酮对子宫内膜细胞凋亡率显著增加(P<0.05),Caspase蛋白表达水平降低。15μg/ml的米非司酮对细胞48小时处理后,与对照组相比,Ang-1和b FGF基因表达水平显著降低(P<0.01),而VEGF与Ang-2基因表达水平明显增加(P<0.05)。结论:米非司酮能诱导人子宫内膜细胞凋亡,促血管生成因子VEGF与Ang-2的高表达以及Ang-1和b FGF的低表达,这可能是米非司酮治疗围绝经期功能失调性子宫出血的重要机制。

Objective： To explore the effects of mifepristone on apoptosis in human endometrial cells and the expression of angiogenic factor Ang 1,Ang 2,VEGF and b FGF. Methods： Using MTS assay to examine the toxicity of mifepristone with different concentrations on endometrial cells,using annexin V / PI staining and flow cytometry technique and Western blot to examine the apoptosis,and using real-time fluorescence quantitative polymerase chain reaction to detect the expression of Pro angiogenic factor. Results： Cytotoxicity of mifepristone increased with increasing time,and decreased with the decreasing drug concentration,IC50 of 24 h,48 h and 72 h is 18 μg / ml,15 μg / ml,12 μg / ml,for respectively. Compared to the control group,the apoptosis of endometrial cells with different concentrations of mifepristone was significantly increased（ P〈0. 05）,and the expression pro-Caspase protein was decreased. With 15 μg / ml mifepristone treated for 48 h,compared to the control group,the expression of Ang-1 and b FGF was significantly decreased（ P〈0. 01）,while VEGF and Ang-2 were significantly increased（ P〈0. 05）. Conclusions： Mifepristone can induce the apoptosis of human endometrial cells,promote the high expression of angiogenesis factor VEGF and Ang-2 and low expression of Ang-1 and b FGF. It may be an important mechanism for mifepristone to treat the perimenopausal dysfunctional uterine bleeding.