不育症患者精浆弹性蛋白酶检测的临床意义

Clinical significance of elastase detection in seminal plasma of infertility patient

目的研究男性不育患者精浆中弹性蛋白酶的含量变化和临床意义,探讨其在诊断无症状性生殖道炎症中的作用;以及其对精子DNA完整性的保护作用的研究。方法收集140例不育症门诊患者的精浆标本(实验组);按WHO标准80例诊断为弱精子症;60例为精液常数正常的不育症男性。所有患者都没有生殖道感染的症状,3个月前无抗菌治疗。采用ELISA双抗体夹心酶联免疫吸附法检测患者精浆弹性蛋白酶水平,20例生育男性作对照。采用硝酸还原酶法检测精浆一氧化氮水平;采用吖啶橙染色,计算单链DNA精子比例,对精子DNA完整性进行评估。结果1、140例不育男性患者(实验组)精浆中的弹性蛋白酶、一氧化氮(NO)浓度、精子单链DNA百分比明显高于生育男性(对照组)(P<0.001)。2、弹性蛋白酶浓度与患者年龄(r=0.255,P<0.01)及精液中白细胞数(r=0.569,P<0.01)有正相关关系。3、弹性蛋白酶浓度与精液量(r=0.537,P<0.01)及精子单链DNA比率(r=0.436,P<0.01)呈负相关关系;与精浆NO浓度无相关关系(r=0.083,P=0.331)。结论精浆中粒细胞弹性蛋白酶检测对诊断无症状性生殖道炎症是一种可靠的筛查试验,另外,弹性蛋白酶可能对精子DNA有保护性作用。

Objective To study the clinical significance and the change of neutrophil elastase concentration in seminal plasma of infertility patient. To investigate the role of elastase for diagnosis and prognosis of silent genital tract inflammation, and a protective role for the elastase towards sperm DNA integrity. Methods 140 seminal plasma samples of out-patient （ study group） with infertility were collected, including 80 cases of asthenospermia men and 60 cases of normozoospermic men. None of the patients had clinical signs or symptoms of a genital tract infection. None had been treated by antibiotics within 3 months before enrolment into the study. All patients had given their informed consent for participation. Elastase concentration in seminal plasma was measured by ELISA method. The control group consisted of 20 fertile men who fathered children spontaneously within the study period. Nitric oxide in seminal plasma was detected by the method of nitrate reductase.The percentage of sperm containing single-stranded DNA was calculated by acridine orange staining to evaluate sperm DNA integrity.Results （1）The prevalence of increased seminal elastase,nitric oxide and he percentage of spermatozoa with single-stranded DNA in 140 infertile men was significantly higher as compared with that observed in 20 fertile men （P 〈 0.01）. （2）A significant positive correlation was found between elastase concentration and patient age （r = 0. 255, P 〈 0.01 ） and the number of leukocytes （r = 0. 569, P 〈 0.01）. （3）A negative correlation was found between elastase concentration and semen volume （ r = - 0. 537, P 〈 0.01 ） and the percentage of spermatozoa with single-stranded DNA （ r = - 0.436, P 〈 0.01 ）, but there was no correlation between elastase and nitrio oxide （ r = 0. 083, P = 0.331 ）. Conclusion Seminal granulocyte elastase is a reliable screening test for silent genital tract inflammation of the couple. Moreover, the elastase may have a protective effect in reducing sperm DNA denaturation.