3类洗涤法处理不同质量精液标本的效果比较及在IUI中的应用选择

Comparison and Selection among Three Common Treatment Methods Processing Different Quality of Semen Specimen

目的:通过比较密度梯度离心法(density-gradient centrifugation,DGC)、上游法(swim-up,SU)和简单洗涤法(simple washing,SW)处理不同质量精液标本的效果,优选出最合适的宫腔内人工授精(Intrauterine insemination,IUI)精液洗涤方法。方法:选取男科实验室行精液常规检查的患者中收集精液量≥3 m L的精液标本90份,其中精子浓度<15×10~6/m L的少精症患者精液标本23份,浓度与活力正常(浓度≥15×10~6/m L,PR≥32%且PR+NP≥40%)的精液标本67份,每份精液标本各取3个1 m L分别用密度梯度离心法、上游法和简单洗涤法处理。通过不同方法的洗涤处理,比较各组精子洗涤前后的浓度、活力(PR)及回收率。结果:3种处理方法洗涤后的PR较洗涤前均有显著提高(P<0.05);在处理后的精子浓度方面,简单洗涤法处理要显著高于其余两类方法(P<0.05);在处理后的PR和活率(PR+NP)方面,密度梯度离心法和上游法又要高于简单洗涤法(P<0.05);在回收率方面,密度梯度离心法和简单洗涤法则显著高于上游法组(P<0.05)。结论:在处理少精症患者精液标本时,上游法因其较低的回收率不适宜被采用;当精子浓度<15×10~6/m L且精子总数小于正常值(39×10~6个)时,为保证处理后最终前向精子总数>10×10~6个,精子标本不适合过多次数的洗涤处理,宜采用简单洗涤法;而只有当精液体积较大时,在保证IUI精子终浓度的同时,密度梯度离心法较简单洗涤法能更好地去除白细胞和不成熟生殖细胞等杂质,而更适宜被采用;在处理浓度活力正常精液标本时,密度梯度离心法和上游法较之简单洗涤法能获得更高的精子活力(P<0.05)而更适宜被采用;同时,密度梯度离心法因较之上游法具有更高的回收率(P<0.05),且具备操作易标准化和操作时间短等特点而更具实用价值。

Objective：To select the most appropriate IUI sperm washing methods through comparative analysis of density-gradient centrifugation,swim-up and simple washing processing different quality of semen specimen.Method：90 semen specimens（ semen volume≥3 m L）from laboratory of andrology were obtained.Among them,23 semen specimens were obtained from oligospermatism patients（sperm concentration15×10^6/m L）,the other 67 semen specimens were obtained from normal men（sperm concentration≥15×10^6/m L,PR≥32%,PR＋NP≥40%）.Each semen sample were taken with 1 m L for processing by density gradient centrifugation,swim-up and simple washing methods respectively.80% and 40% pure ception were used in density-gradient centrifugation.Sperm concentration,motility and recovery rate were compared before and after treatment by the three methods.Result：Sperm motility（PR） increased obviously after treated by all the three methods（P〈0.05）.Sperm concentration after treated by simple washing was significantly higher than other two methods（P〈0.05）.PR after treated by density-gradient centrifugation and swim-up were significantly higher than simple washing（P〈0.05）.Sperm recovery rate after treated by density-gradient centrifugation and simple washing were significantly higher than swim-up（P〈0.05）.Conclusion：Swim-up is not suitable for oligospermatism patients for its low sperm recovery rate.When sperm concentration〈15×10^6/m L and total semen count39×10^6,we choose simple washing to ensure that final sperm count〉10×10^6 and avoid frequently washing.When semen volume is large enough,for density-gradient centrifugation can reduce leukocyte and immaturity cell better than simple washing,it is more appropriate be selected.When PR and concentration are normal,density-gradient centrifugation and swim-up are both better than simple washing.However,for sperm recovery rate of densitygradient centrifugation is higher than swim-up,and it is easier be standardized and cost less time for operation,the density-gradient centrifugation is the more appropriate method for IUI.