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目的:探讨体外受精-胚胎移植(IVF-ET)中精子处理前、后凋亡率的变化以及处理前、后精子凋亡率与受精率和胚胎质量的相关性。方法:收集行IVF者的精液55例,密度梯度离心结合上游法处理精液,IVF当日通过流式细胞术检测处理前、后精子凋亡指标——caspase-3(CP3)活化率,于受精后16 h、48 h、72 h分别观察受精率、卵裂率、优质胚胎率。结果:共获卵690枚,平均受精率、卵裂率、优质胚胎率分别为:84.49±14.92%、94.62±11.09%、47.11±27.17%;处理后精子CP3活化率较处理前有明显降低(22.84±6.05%vs 34.80±6.56%,P<0.05);处理前精子CP3活化率与受精率、卵裂率、优质胚胎率均无显著相关性(Spearman相关系数:0.056,0137,-0.060,P>0.05),处理后精子CP3活化率与受精率及优质胚胎率呈显著负相关(Spearman相关系数:-0.375,-0.416,P<0.01),与卵裂率无显著相关性(Spearman相关系数:-0.156,P>0.05)。结论:密度梯度离心结合上游法处理精子后可以显著降低精子凋亡率,处理后精子凋亡率可能影响受精率和优质胚胎率,可为临床预测受精结局及胚胎发育情况提供参考指标。
Objective: To investigate the changes of apoptosis rate before and after spermatozoa in vitro fertilization and embryo transfer (IVF-ET) and the correlation between sperm apoptosis rate and fertilization rate and embryo quality before and after treatment. Methods: Fifty-five semen of IVF were collected, and semen was collected by density gradient centrifugation combined with the upstream method. The activation of caspase-3 (CP3), a marker of sperm apoptosis before and after treatment, was detected by flow cytometry on day IVF. 16 h, 48 h, 72 h were observed fertilization rate, cleavage rate, high-quality embryo rate. Results: The average fertilization rate, cleavage rate and high quality embryos were 84.49 ± 14.92%, 94.62 ± 11.09% and 47.11 ± 27.17%, respectively. The activated rate of CP3 was significantly lower than that before treatment 22.84 ± 6.05% vs 34.80 ± 6.56%, P <0.05). There was no significant correlation between CP3 activation rate and fertilization rate, cleavage rate and good embryo rate before treatment (Spearman correlation coefficient: 0.056,0137,0.060, P > 0.05). There was a significant negative correlation between sperm CP3 activation rate and fertilization rate and high quality embryo rate (Spearman correlation coefficient: -0.375, -0.416, P <0.01), but not cleavage rate (Spearman correlation coefficient: -0.156, P> 0.05). Conclusions: Density gradient centrifugation combined with upstream treatment of spermatozoa can significantly reduce the rate of sperm apoptosis. After treatment, the rate of sperm apoptosis may affect the fertilization rate and the rate of high quality embryos, which may provide a reference index for clinical prediction of fertilization outcome and embryo development.