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目的本研究探讨不同浓度二甲双胍在不同时间点对其分泌类固醇激素的影响。方法收集因输卵管因素不孕或者男性因素不孕行IVF/ICSI-ET患者促排卵后的含有黄素化颗粒细胞的卵泡液,经密度梯度离心后分离获得颗粒细胞,行体外培养。分别加入含有二甲双胍浓度为0 ng/ml,10 ng/ml,100ng/ml,1000ng/ml,10000ng/ml的DMEM培养基对其进行体外培养,每24h收集细胞培养上清,用化学发光法检测上清液中雌二醇及孕酮的水平。结果体外培养颗粒细胞分泌类固醇激素在72h到96h达到高峰,120h后开始递减,这与二甲双胍体外生长曲线一致;二甲双胍都能够抑制类固醇激素的分泌,且二甲双胍浓度在10000ng/ml,能够极显著抑制类固醇激素的分泌,在每个时间点均呈剂量依赖性。结论二甲双胍在颗粒细胞体外发育整个过程中均抑制。
Objectives This study was to investigate the effects of different concentrations of metformin on the secretion of steroid hormones at different time points. Methods The follicular fluid containing luteinized granulosa cells after IVF / ICSI-ET infertility due to tubal infertility or male infertility was collected, and granulosa cells were isolated by density gradient centrifugation and cultured in vitro. The cells were cultured in DMEM containing Metformin at a concentration of 0 ng / ml, 10 ng / ml, 100 ng / ml, 1000 ng / ml, and 10000 ng / ml, respectively, and the cell culture supernatants were harvested every 24h. Chemiluminescence Estradiol and progesterone levels in supernatant. RESULTS: Secretion of steroid hormones by granulosa cells in vitro reached a peak at 72h to 96h and began to decrease after 120h, which was consistent with the growth curve of metformin in vitro. Metformin inhibited the secretion of steroid hormones and the concentration of metformin at 10000ng / ml significantly inhibited steroid Hormone secretion was dose-dependent at each time point. Conclusion Metformin inhibits the growth of granulosa cells in vitro.