GnRH拮抗剂联合来曲唑和米非司酮对体外培养的人颗粒细胞功能的影响

目的探讨促性腺激素释放激素拮抗剂(GnRH-ant)联合来曲唑和米非司酮对体外培养的人卵巢颗粒细胞功能及血管内皮生长因子(VEGF)表达的影响。方法体外培养人原代卵巢颗粒细胞及人卵巢颗粒细胞系KGN,根据所用药物不同分为GnRH-ant、来曲唑、米非司酮以及上述三种药物联合用药组,药物干预后检测细胞活力、细胞凋亡、培养液中雌二醇(E 2)及孕酮(P)水平及VEGF的表达。结果(1)GnRH-ant、来曲唑、米非司酮三种药物均可以不同程度地抑制原代颗粒细胞和KGN细胞的活力、促进颗粒细胞凋亡,其中联合用药组的作用最强(P<0.05);(2)GnRH-ant、来曲唑、米非司酮三种药物均可以不同程度地抑制原代颗粒细胞和KGN细胞E 2、P分泌水平,其中联合用药组E 2、P水平最低且差异有统计学意义(P<0.05);(3)GnRH-ant、来曲唑、米非司酮三种药物均可以不同程度地抑制原代颗粒细胞和KGN细胞VEGF表达,其中联合用药组VEGF mRNA及蛋白表达水平最低(P<0.05)。结论GnRH-ant联合来曲唑和米非司酮可以有效地抑制颗粒细胞活力、减少类固醇激素分泌及降低VEGF的表达水平。

Effects of GnRH antagonist combined with letrozole and mifepristone on function of human ovarian granulosa cell in vitro

Objective:To investigate the effects of GnRH antagonist(GnRH-ant)combined with letrozole and mifepristone on the function of granulosa cell and the expression of vascular endothelial growth factor(VEGF)in vitro.Methods:Human primary ovarian granulosa cells and human ovarian granulosa cell line KGN were cultured in vitro.According to the different drugs used,the cultured cells were divided to 4 groups:GnRH-ant,letrozole,mifepristone,and combination of the three drugs groups;the blank treatment was used as control group.Cell viability,cell apoptosis,the levels of estradiol(E 2)&progesterone(P)and VEGF were also detected after treatment.Results:GnRH-ant,letrozole and mifepristone inhibited the cell viability,and promoted the apoptosis of primary granulosa cells and KGN cell compared with the control group,and the combination group of the three drugs had the strongest effect(P<0.05).The E 2 and P levels of primary granulosa cells and KGN cells in GnRH-ant,letrozole,mifepristone and the combination group were significantly lower than those in the control group respectively,and the levels of the combination group were the lowest(P<0.05).GnRH-ant,letrozole,and mifepristone inhibited the expression of VEGF in primary granulosa cells and KGN cells to varying degrees,and the expression of VEGF mRNA and protein in the combination group was the lowest(P<0.05).Conclusions:GnRH-ant combined with letrozole and mifepristone can effectively inhibit cell viability,promote cell apoptosis,reduce the secretion of steroid hormones and decrease the express level of VEGF of ovarian granulosa cells.