促排卵期间精子DNA碎片指数和线粒体膜电位联合检测对辅助生殖技术妊娠结局的预测分析

Predictive analysis of combined detection of sperm DNA fragmentation index and mitochondrial membrane potential during ovulation induction on assisted reproductive technology pregnancy outcomes

目的探讨促排卵期间精子DNA碎片指数(DNA fragmentation index,DFI)和线粒体膜电位(mitochondrial membrane potential,MMP)联合检测对辅助生殖技术(assisted reproductive technology,ART)妊娠结局的预测作用。方法2020年4月至2021年5月,在海军军医大学第一附属医院生殖医学中心行体外受精或卵细胞质内单精子注射治疗的患者中,在女方促排卵期间接受精子DFI和(或)MMP检测并符合纳入标准的病例共116个周期。最近一次检测时间大多安排在促排卵起始日、夜针日或取卵日。采用流式细胞仪检测荧光来计算精子DFI值与MMP值。DFI值≤15%为正常值组,15%~30%为临界值组,≥30%为异常值组;DFI正常值和临界值的所有病例均检测MMP值,MMP值<30%为MMP异常值组。比较各组间的优质胚胎率、胚胎种植率和中晚期妊娠率。统计学方法采用t检验、方差分析、χ^(2)检验、非参数检验和二元Logistic回归分析。结果DFI正常值组、临界值组与异常值组3组间男方年龄、女方年龄和不育年限比较,差异无统计学意义(P>0.05)。DFI正常值组、临界值组与异常值组的取卵周期分别为44、33和39个,3组间比较优质胚胎率[54.4%(198/364)、56.5%(130/230)与39.5%(83/210),χ^(2)=15.550,P<0.01]、胚胎种植率[48.9%(44/90)、42.4%(25/59)与25.0%(15/60),χ^(2)=8.709,P<0.05]和中晚期妊娠率[70.6%(36/51)、44.4%(16/36)与30.6%(11/36),χ^(2)=14.471,P<0.01],差异均有统计学意义。进一步两两比较,DFI正常值组的优质胚胎率、胚胎种植率和中晚期妊娠率均高于DFI异常值组;DFI正常值组的中晚期妊娠率高于DFI临界值组;DFI临界值组的优质胚胎率和胚胎种植率均高于DFI异常值组。MMP异常值组和正常值组的取卵周期分别为26和51个,MMP异常值组与正常值组的优质胚胎率[46.0%(57/124)与57.7%(271/470),χ^(2)=5.424,P<0.05]、胚胎种植率[18.2%(8/44)与58.1%(61/105),χ^(2)=19.867,P<0.01]和中晚期妊娠率[24.0%(6/25)与74.2%(46/62),χ^(2)=18.667,P<0.01]比较,MMP异常值组均低于正常值组。采用多因素Logistic回归分析矫正混杂因素后,结果提示,DFI值(OR=0.892,95%CI:0.801~0.995,P<0.05)和MMP值(OR=1.068,95%CI:1.009~1.132,P<0.01)是影响中晚期妊娠率的独立影响因素,DFI值越高,中晚期妊娠率越低;MMP值越低,中晚期妊娠率越低。结论在女方促排期间检测的精子DFI值与优质胚胎率、胚胎种植率和中晚期妊娠率有显著相关性;DFI值正常或临界的患者中,MMP异常也会显著降低胚胎种植率和中晚期妊娠率;DFI值和MMP值两者联合检测可预测ART妊娠结局。

Objective To explore the predictive effect of sperm DNA fragmentation index(DFI)and mitochondrial membrane potential(MMP)combined detection on assisted reproductive technology(ART)pregnancy outcomes during ovulation induction.Method A total of 116 in vitro fertilization/intracytoplasmic sperm injection cycles which men underwent sperm DFI and/or MMP detection during ovulation induction were enrolled at the Reproductive Center of the First Affiliated Hospital of Naval Medical University between April 2020 and May 2021.The last testing times were scheduled on the start day of ovulation induction,night needle day or egg retrieval day.Fluorescence was detected by flow cytometry to calculate sperm DFI and MMP value.Patients with DFI≤15%,15%~30%and≥30%were normal value group,critical value group and abnormal value group,respectively.MMP value was detected in all cases with normal DFI value and critical value.Patients with MMP values<30%were abnormal MMP value group.The high-quality embryo rate,embryo implantation rate and mid-to-late pregnancy rate were compared among the groups.Nonparametric test,t-test,variance analysis,χ^(2) test and binary Logistic regression analysis were used for statistical analysis.Result There was no significant difference in the age of the man,the age of the woman and the years of infertility among the three groups of DFI normal value group,critical value group and abnormal value group(P>0.05).DFI normal value group,critical value group and abnormal value group had 44,33 and 39 retrieval cycles,respectively.There were statistically significant differences in the high-quality embryo rate[54.4%(198/364),56.5%(130/230)and 39.5%(83/210),χ^(2)=15.550,P<0.01],embryo implantation rate[48.9%(44/90),42.4%(25/59)and 25.0%(15/60),χ^(2)=8.709,P<0.05]and mid-to-late pregnancy rate[70.6%(36/51),44.4%(16/36)and 30.6%(11/36),χ^(2)=14.471,P<0.01]among the three groups.Further pairwise comparisons showed that the high-quality embryo rate,embryo implantation rate and mid-to-late pregnancy rate in the normal DFI value group were higher than those in the abnormal DFI value group;the mid-to-late pregnancy rate in the normal DFI value group was higher than that in the critical DFI value group;the high-quality embryo rate and embryo implantation rate in the critical DFI value group were higher than those in the abnormal DFI value group.The oocyte retrieval cycles of the abnormal MMP value group and the normal value group were 26 and 51,respectively.The high-quality embryo rate[46.0%(57/124)vs 57.7%(271/470),χ^(2)=5.424,P<0.05],embryo implantation rate[18.2%(8/44)vs 58.1%(61/105),χ^(2)=19.867,P<0.01],and mid-to-late pregnancy rate[24.0%(6/25)vs 74.2%(46/62),χ^(2)=18.667,P<0.01]of the abnormal MMP group were lower than those of the normal group.After adjusting for confounding factors by multivariate Logistic regression analysis,the results showed that the DFI value(OR=0.892,95%CI:0.801-0.995,P<0.05)and the MMP value(OR=1.068,95%CI:1.009-1.132,P<0.01)were independent factors affecting the mid-to-late pregnancy rate.The higher the DFI value,the lower the mid-to-late pregnancy rate;the lower the MMP value,the lower the mid-to-late pregnancy rate.Conclusion The DFI value of sperm detected during the ovulation induction period of the woman's ovulation has a significant correlation with the rate of high-quality embryo rate,embryo implantation rate and mid-to-late pregnancy rate.Abnormal MMP in the normal and critical DFI patients significantly reduces the embryo implantation rate and mid-to-late pregnancy.The combined detection of DFI and MMP can predict the outcomes of ART pregnancy.