ICSI治疗患者卵丘颗粒细胞自噬活性对卵母细胞质量的影响

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目的:探讨卵泡浆内单精子注射(ICSI)治疗患者卵丘颗粒细胞自噬活性及凋亡水平对卵母细胞质量的影响。方法:对行ICSI治疗的50例患者常规促排卵后获得220个卵子采用透明质酸酶处理,获取卵丘细胞,并对卵子成熟度进行评估。选取成熟的MII期卵子行ICSI操作,采用TUNEL试验检测细胞凋亡情况,免疫荧光检测卵丘颗粒细胞自噬活性。结果:GV期卵子荧光强度高于MI期及MII期,LC3-II/I比值低于MI期及MII期,MI期子荧光强度高于MII期,LC3-II/I比值低于MII期,两两比较差异有统计学意义(P<0.05)。受精组卵子荧光强度低于未受精组,LC3-II/I高于未受精组,差异有统计学意义(P<0.05)。随着Dale胚胎评分的增加,卵子荧光强度显著减少,而LC3-II/I显著提高,两两比较差异有统计学意义(P<0.05)。与流产组相比,活产组卵子荧光强度显著减少,而LC3-II/I显著提高,差异有统计学意义(P<0.05)。结论:卵丘颗粒细胞自噬活性及凋亡水平可影响卵子的成熟度、质量、受精、胚胎发育及妊娠结局。 Objective: To investigate the effect of intracytoplasmic sperm injection (ICSI) on the granulosa cell autophagy activity and the level of apoptosis on oocyte quality. Methods: Fifty patients undergoing ICSI treatment received 220 eggs after routine ovulation induction. Hyaluronidase (Hyaluronidase) treatment was used to obtain cumulus cells and the maturity of the eggs was evaluated. The mature MII oocytes were selected for ICSI. TUNEL assay was used to detect apoptosis. Immunofluorescence was used to detect the autophagy activity of cumulus granule cells. Results: The fluorescence intensity of GV ova was higher than that of MI and MII, the ratio of LC3-II / I was lower than MI and MII, the fluorescence intensity of MI was higher than that of MII and the ratio of LC3-II / I was lower than MII, The difference was statistically significant (P <0.05). The egg fluorescence intensity of the fertilized group was lower than that of the unfertilized group, and the LC3-II / I was higher than that of the unfertilized group (P <0.05). With the increase of Dale embryo score, the egg fluorescence intensity was significantly reduced, while the LC3-II / I significantly increased, the difference was statistically significant (P <0.05). Compared with the abortion group, the ovum fluorescence intensity of live births significantly decreased, while LC3-II / I significantly increased, the difference was statistically significant (P <0.05). Conclusion: The autophagy activity and apoptosis of granulosa cells may affect the maturation, quality, fertilization, embryo development and pregnancy outcome.
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