密度梯度离心后精子悬液中弹性硬蛋白酶浓度和精子DNA完整性的测定

The influence of density gradient centrifugation technique to seminal PMN-Elastase and sperm DNA integrity

目的检测体外授精过程中精液和经密度梯度离心后精子悬液中弹性硬蛋白酶(PMN-Elastase)的浓度与精子DNA的完整性,探讨密度梯度离心对精浆中PMN-Elastase的作用和精子DNA的损伤情况。方法选择我中心64例进入体外受精周期患者,其中根据女方取卵手术当天男方精液检测PMN—Elastase不同浓度将患者分为A组(15例):含量>1000ng/ml(显性感染);B组(20例):含量290-1000ng/ml(隐性感染);C组(20):含量<290ng/ml(正常水平),分析和比较处理前后三组精子悬液中PMN-Elastase的浓度和DNA完整率。结果三组处理后的精子悬液中A组PMN-Elastase前后浓度没有明显区别(P>0.05),B组和C组浓度明显下降(P<0.05);三组处理后的DNA完整率均明显提高(P<0.05),经过长时间孵育均明显降低(P<0.05)。结论高浓度PMN-Elastase的精液经密度梯度离心后的精子悬液中PMN-Elastase浓度并不增加,精子活力和DNA完整率增加,可能不影响受精结局。

Objective：To detect the level of PMN-Elastase in semen after the density gradient centrifugation and spermatozoa DNA damage during in vitro fertilization（IVF）,and discuss the effect of density gradient centrifugation to PMN-Elastase concentration and DNA damage. Method：This study involved 64 IVF cycles,on the day of ovum pick-up the male patients sperm were detected included PMN-Elastase and sperm DNA integrity. on the basis of PMN-Elastase levels,the samples were divided into A group（1000ng/ml）consisted of 15 cases,B group（290-1000ng/ml）20 cases and C group（290ng/ml）20cases. Then compare the difference in three groups.Results：After the density gradient centrifugation,the outcomes of seminal PMN-Elastase between A and B group were decreased obviously,and after treatment the DNA integrity of three groups all increased significantly and a long incubation about 18 hours degraded. Conclusion：After the density gradient centrifugation,it does′t increase the concentration of PMN-Elastase（the apparent infection group）in sperm suspension in the seminal plasma infection patients and increase the sperm DNA integrity,may not influence the fertilization outcome.