GnRH激动剂对子宫内膜异位症患者卵巢颗粒细胞细胞周期及抗凋亡蛋白cFLIP表达的影响

目的探讨GnRH激动剂（GnRHa）治疗对子宫内膜异位症（EMs）患者卵巢颗粒细胞细胞周期以及抗凋亡蛋白cFLIP表达的影响。方法共45例接受体外授精-胚胎移植（IVF-ET）治疗的患者,其中EMs患者19例,随机分配至GnRH-a治疗组（A组,n=11）和常规长方案治疗组（B组,n=8例）,另26例管性因素不孕的常规长方案治疗患者为对照组（C组）。各组取卵时收集卵泡液颗粒细胞,采用流式细胞仪检测细胞周期,Western blotting测定颗粒细胞cFLIP蛋白含量。结果三组的受精率、临床妊娠率、种植率及流产率差异均无统计学意义（P〉0.05）。三组患者G0/G1期细胞比例的差异无统计学意义（P〉0.05）;A组和B组S期细胞比例显著高于C组,G2/M期细胞比例显著低于C组（P〈0.05）,凋亡峰显著高于C组（P〈0.05）;但A组与B组细胞周期各时相差异均无统计学意义（P〉0.05）。C组卵巢颗粒细胞cFLIP蛋白表达水平显著高于A和B组（P〈0.05）,A、B两组cFLIP蛋白表达差异无统计学意义（P〉0.05）。结论 EMs患者卵巢颗粒细胞凋亡升高,且细胞周期异常;延长的GnRH-a治疗并不改善这种异常以及抗凋亡蛋白cFLIP的低表达状态。

Effects of GnRH agonist on cell cycle and expression of anti-apoptosis protein cFLIP of ovarian granulosa cells in patients with endometriosis

Objective To investigate the effects of GnRH agonist (GnRH-a) on cell cycle and expression of anti-apoptosis protein cFLIP of ovarian granulosa cells in patients with endometriosis. Methods Forty-five women treated with in vitro fertilization and embryo transfer (IVF-ET) were selected, among whom 19 were patients with endometriosis and randomly assigned to GnRH-a treatment group (group A, n=11) and general long protocol group (group B, n=8). The other 26 women who were infertile due to tubal factors and treated with general long protocol were served as control group(group C). Granulosa cells were obtained from follicular fluid, cell cycle was analyzed by flow cytometry, and the expression of cFLIP protein in granulosa cells was detected by Western blotting. Results There was no significant difference in fertilization rates, clinical pregnancy rates, implantation rates and miscarriage rates among three groups (P>0.05). There was no significant difference in the percentage of G0/G1 phase cells among three groups (P>0.05). The percentage of S phase cells was significantly higher, that of G2/M phase cells was significantly lower, and that of apoptosis peak was significantly higher in group A and group B than those in group C (P<0.05). However, there was no significant difference in cell cycle between group A and goup B (P>0.05). The expression of cFLIP protein in ovarian granulosa cells in group C was significantly higher than that in group A and group B (P<0.05), while there was no significant difference between group A and group B (P>0.05). Conclusion The apoptosis of ovarian granulosa cells in patients with endometriosis increases, with abnormal cell cycle. Treatment with prolonged GnRH-a protocol may not improve the abnormal status of impaired cell cycle and lower expression of cFLIP.