应用单层和双层密度梯度离心法处理男性不育症患者精液效果对比

Comparison of application of single-layer and double-layered gradient centrifugation in artificial insemination with husband′s semen

目的评价单层和双层密度梯度离心法处理男性不育症患者精液并行宫腔内人工授精(artificial insemination with husband′s sperm,AIH IUI)的应用效果。方法连续收集在本中心施行IUI手术的男方精液标本500份,随机采用70%单层法(正常精子45份、轻度少精子症90份、轻度弱精子症90份)或90%+45%双层法(正常精子55份、轻度少精子症110份、轻度弱精子症110份)进行密度梯度离心,优选前后均进行精子质量和功能检测,并随访妊娠结局。结果经单层法优选后,正常精子组前向运动精子百分率(PR)和PR精子回收率低于双层法,轻度少精子症组精子总活力[PR+非前向运动镜子百分率(NP)]和PR+NP精子回收率高于双层法(均P<0.05);轻度弱精子症组经两种方法优选后,PR和PR+NP精子回收率高于正常精子组和轻度少精子症组(P<0.05);3组精液经两种方法优选后,正常形态精子百分率、诱发顶体反应(AR)率、DNA碎片化指数(DFI)、精子成熟度和精液白细胞浓度与处理前差异均有统计学意义(P<0.05);研究时间段内采用来曲唑(LE)促排方案的334个周期临床妊娠率,单层法68周期,正常精子15.38%,轻度少精子症14.81%,轻度弱精子症17.86%;双层法266周期,正常精子13.21%,轻度少精子症14.15%,轻度弱精子症15.89%(均P>0.05)。结论单层和双层密度梯度离心法均能筛选较高质量的精子,应根据不同病因患者的精液情况选用合适的优选方法。

Objective To evaluate the effect of single layer and two layer density gradient centrifugation in artificial insemination(AIH IUI).Methods 500 semen specimens from normozoospermia,mild oligozoosper mia and mild asthenozoospermia men were collected at our clinic.After 70%single layer(n=225)and 90%+45%two layer(n=275)density gradient centrifugation,the concentration,motility,progressive motility and func tion test of sperm were respectively analyzed.Results After sperm was separated by the single layer gradient,the percentage of progressive motility(PR)sperm and the recovery rate of PR sperm in normozoospermia group were significant lower than that by the two layer gradient,and the motility(PR+NP)sperm and the recovery rate of PR+NP sperm in the oligospermia group were significant higher than that by the two layer gradient(P<0.05).The recovery rate of PR and PR+NP sperm in asthenospermia group was significant higher than that in normozoosper mia group and asthenospermia group(P<0.05).The normal morphology of sperm,acrosome reaction,DNA integ rity,maturity and leukocyte concentration of the single and two layer gradient were significantly better than pre separation(P<0.05),but there were no significantly difference between two methods(P>0.05).There were 334 letrozole(LE)stimulate ovulation IUI cycles in these specimens,the clinical pregnancy rate of 68 single layer cycles was 16.18%(normozoospermia group is 15.38%,oligozoospermia group is 14.81%and group asthenozoo spermia is 17.86%,respectively).The clinical pregnancy rate of 266 double layer cycles was 14.66%(normozoo spermia group is 13.21%,oligozoospermia group is 14.15%and asthenozoospermia group is 15.89%,respectively).There were no significantly difference among three groups and two methods(P>0.05).Conclusion The high quality sperm can be screened by double layer and monolayer density gradient centrifugation,and appropriate selection method should be chosen according to the semen situation of the patients with different etiology.