精子线粒体早老素相关菱形样蛋白及精子DNA碎片指数与体外受精-胚胎移植妊娠结局的相关性分析

Correlations of sperm mitochondrial presenilin associated rhomboid like protein and sperm DNA fragmentation index with pregnancy outcome of in vitro fertilization embryo transfer

目的观察行体外受精-胚胎移植的不孕夫妇中男性患者精子DNA碎片指数(DNA fragmentation index, DFI)和精子线粒体早老素相关菱形样蛋白(presenilin associated rhomboid like protein, PARL)水平变化,探讨其与体外受精-胚胎移植妊娠结局的关系。方法行体外受精-胚胎移植的不孕夫妇中男性患者140例,根据临床妊娠情况分为妊娠成功组57例和妊娠失败组83例。2组行体外受精-胚胎移植前,采用精子染色质扩散法检测精子DFI,采用ELISA法检测精子线粒体PARL水平,比较2组年龄、精子密度、精子活力、精子DFI、精子线粒体PARL水平及优质胚胎率等临床资料。以精子DFI、精子线粒体PARL水平的平均值25%、5.86μg/L为临界值,将140例患者分为高DFI组62例(DFI≥25%)和低DFI组78例(DFI<25%),高PARL组71例(PARL≥5.86μg/L)和低PARL组69例(PARL<5.86μg/L);比较高、低DFI、PARL组精子密度、精子活力、精子畸形率、优质胚胎率、种植率、卵裂率、受精率、临床妊娠率;采用Pearson或Spearman相关性分析精子线粒体PARL水平、精子DFI分别与精子密度、精子活力、优质胚胎率、临床妊娠率、精子畸形率的相关性;采用多因素logistic回归分析体外受精-胚胎移植妊娠失败的影响因素。结果妊娠成功组优质胚胎率(47.30%)、精子密度[(27.42±4.13)×10^(6)/mL]、精子活力[(51.84±13.02)%]、精子线粒体PARL水平[(13.54±2.01)μg/L]均高于妊娠失败组[31.82%、(22.09±3.26)×10^(6)/mL、(32.28±8.12)%、(4.51±1.07)μg/L](P<0.05),精子畸形率[(16.80±4.82)%]、精子DFI[(10.98±3.06)%]均低于妊娠失败组[(26.83±6.94)%、(30.46±6.18)%](P<0.05)。高DFI组优质胚胎率(30.00%)、临床妊娠率(30.65%)、精子密度[(20.14±4.36)×10^(6)/mL]、精子活力[(24.45±8.73)%]均低于低DFI组[46.89%、48.72%、(27.54±4.12)×10^(6)/mL、(52.81±13.04)%](P<0.05),精子畸形率[(30.12±7.93)%]高于低DFI组[(16.87±5.94)%](P<0.05);高PARL组优质胚胎率(47.96%)、临床妊娠率(53.52%)、精子密度[(27.25±4.12)×10^(6)/mL]、精子活力[(50.66±12.95)%]均高于低PARL组[28.68%、27.54%、(21.04±4.23)×10^(6)/mL、(24.76±8.87)%](P<0.05),精子畸形率[(16.72±5.63)%]低于低PARL组[(30.75±7.81)%](P<0.05);精子线粒体PARL与精子密度(r=0.507,P=0.007)、精子活力(r=0.514,P=0.009)、优质胚胎率(r=0.470,P<0.001)、临床妊娠率(r=0.564,P=0.002)均呈正相关,与精子畸形率呈负相关(r=-0.489,P=0.007);精子DFI与精子密度(r=-0.496,P=0.008)、精子活力(r=-0.503,P=0.006)、优质胚胎率(r=-0.520,P=0.006)、临床妊娠率(r=-0.483,P=0.031)呈均负相关,与精子畸形率呈正相关(r=0.512,P=0.004)。精子线粒体PARL(OR=0.842,95%CI:0.759~0.934,P=0.001)、精子DFI(OR=2.426,95%CI:1.334~4.411,P=0.004)是体外受精-胚胎移植后妊娠失败的影响因素。结论体外受精-胚胎移植妊娠失败的不孕夫妇中男性患者精子DFI升高、精子线粒体PARL水平降低,二者是体外受精-胚胎移植妊娠失败的影响因素。

Objective To observe the levels of sperm mitochondrial presenilin associated rhomboid like protein(PARL)and sperm DNA fragmentation index (DFI)in infertile male patients whose wives received in vitro fertilization embryo transfer(IVF-ET),and to investigate their correlations with the pregnancy outcome.Methods A total of 140infertile male patients whose wives received IVF-ET were divided into successful pregnancy group(n=57)and failed pregnancy group(n=83)according to clinical pregnancy outcome.The levels of DFI in sperm and PARL in sperm mitochondria were detected by sperm chromatin dispersion method and ELISA technique before IVF-ET.The age,sperm density,sperm motility,sperm DFI and PARL in sperm mitochondria,and high-quality embryo rate were compared between two groups.Taking the mean values of sperm DFI and PARL (25%,5.86μg/L)as the cut-off values,140 males were divided into 62males with DFI≥25%(high DFI group)and 78males with DFI<25%(low DFI group),as well as 71males with PARL≥5.86μg/L (high PARL group)and 69males with PARL<5.86μg/L (low PARL group).The sperm density,sperm motility,sperm deformity rate,high-quality embryo rate,implantation rate,cleavage rate,fertilization rate,and pregnancy rate were compared between high and low DFI groups and between high and low PARL groups.Pearson or Spearman correlation analysis was done to assess the correlations of PRAL and DFI with sperm density,sperm motility,high-quality embryo rate,pregnancy rate and sperm deformity rate.Multivariate logistic regression analysis was done to assess the influencing factors of IVF-ET failure.Results The high-quality embryo rate(47.30%),sperm density[(27.42±4.13)×10^(6)/mL],sperm motility[(51.84±13.02)%]and PARL level[(13.54±2.01)μg/L]in successful pregnancy group were higher than those in failed pregnancy group[31.82%,(22.09±3.26)×10^(6)/mL,(32.28±8.12)%,(4.51±1.07)μg/L](P<0.05),and the sperm deformity rate[(16.80±4.82)%]and DFI[(10.98±3.06)%]were lower than those in failed pregnancy group[(26.83±6.94)%,(30.46±6.18)%](P<0.05).The high-quality embryo rate (30.00%),clinical pregnancy rate (30.65%),sperm density[(20.14±4.36)×10^(6)/mL]and sperm motility[(24.45±8.73)%]in high DFI group were lower than those in low DFI group[46.89%,48.72%,(27.54±4.12)×10^(6)/mL,(52.81±13.04)%](P<0.05),and the sperm deformity rate[(30.12±7.93)%]was higher than that in low DFI group[(16.87±5.94)%](P<0.05).The high-quality embryo rate(47.96%),clinical pregnancy rate(53.52%),sperm density[(27.25±4.12)×10^(6)/mL]and sperm motility[(50.66±12.95)%]in high PARL group were higher than those in low PARL group[28.68%,27.54%,(21.04±4.23)×10^(6)/mL,(24.76±8.87)%](P<0.05),and the sperm deformity rate[(16.72±5.63)%]was lower than that in low PARL group[(30.75±7.81)%](P<0.05).The sperm mitochondrial PARL was positively correlated with the sperm density(r=0.507,P=0.007),sperm motility(r=0.514,P=0.009),high-quality embryo rate(r=0.470,P<0.001)and clinical pregnancy rate(r=0.564,P=0.002),and negatively correlated with the sperm deformity rate(r=-0.489,P=0.007).The sperm DFI was negatively correlated with the sperm density(r=-0.496,P=0.008),sperm motility(r=-0.503,P=0.006),high-quality embryo rate(r=-0.520,P=0.006)and pregnancy rate(r=-0.483,P=0.031),and positively correlated with the sperm deformity rate(r=0.512,P=0.004).The sperm mitochondrial PARL (OR=0.842,95%CI:0.759-0.934,P=0.001)and sperm DFI(OR=2.426,95%CI:1.334-4.411,P=0.004)were the influencing factors of pregnancy failure after IVF-ET.Conclusion In those with failed IVF-ET,the sperm DFI increases,and the sperm mitochondrial PARL decreases,both of which are the influencing factors of IVF-ET failure.