男性不育病人精液中磷脂酶A2的测定及其临床意义

目的 :探讨人精液中磷脂酶A2 (PLA2 )含量测定在男性不育症中的临床意义。　方法 :以自制的 2株PLA2单克隆抗体建立酶联免疫吸附试验 (ELISA)、免疫细胞化学法 (ICC)和流式细胞术 (FCM)等 3种方法 ,分别检测男性不育病人精浆及精子头部PLA2的含量 ,并与生育组进行比较。精液常规分析采用计算机辅助精液分析系统 (CASA)进行。　结果 :男性不育病人精浆中PLA2含量分别为 :无精子症组 (31.13± 14 .4 9)ng ml,少精子症组 (17.71±12 .4 5 )ng ml,精子数目正常组 (16 .4 6± 11.31)ng ml;与生育组 (8.0 9± 3.15 )ng ml]相比差异均有极显著性 (P <0 .0 1) ;精浆中PLA2含量与精子密度呈显著负相关 (r=- 0 .6 0 2 ,P <0 .0 5 ) ,而与精子活动力及活率无显著相关性 (r=0 .2 6 6和r=- 0 .2 0 0 ,P均 >0 .0 5 ) ;ICC和FCM试验均提示 ,生育组精子头部PLA2含量显著高于各不育组病人 ,且FCM试验显示差异有极显著性 (P <0 .0 1)。　结论 :精浆中PLA2与男性生育密切相关 ,精子头部PLA2含量缺乏可能是引起男性不育的病因之一。精液中PLA2含量测定方法的建立可为探讨男性不育的发病机制提供有力的依据。

Detection and Clinical Significance of Phospholipase A2 in Semen of Male Infertile Patients

OBJECTIVES: To explore the clinical application of anti-human seminal plasma phospholipase A2 (PLA2) monoclonal antibody (McAb) for male infertility. METHODS: Enzyme-linked immunoabsorbent assay (ELISA), immunocytochemistry(ICC), as well as flow cytometry (FCM) analysis were established using two strains anti-human seminal plasma PLA2 McAb prepared by our laboratory to detect the PLA2 content in human seminal plasma and the anterior head region of spermatozoa, respectively. Then the PLA2 content in male infertile patients were compared with that in normal control with fertility. The seminal routine analysis was performed by computer-assisted semen analysis (CASA). RESULTS: The PLA2 content of infertile groups were (31.13 +/- 14.49) ng/ml in azoospermic patients, (17.71 +/- 12.45) ng/ml in oligospermic patients and (16.46 +/- 11.31) ng/ml in patients with normal sperm density, which were all higher than that of normal controls (8.09 +/- 3.15) ng/ml, P < 0.01]; There was significantly negative correlation between PLA2 content in seminal plasma and sperm density(r = -0.602, P < 0.05), while there was insignificant correlation between PLA2 and sperm motility or percentage of motility. The PLA2 content in the anterior head region of spermatozoon of male infertile groups was significantly lower than that of normal controls by ICC and FCM(P < 0.01). CONCLUSIONS: PLA2 in human seminal plasma is closely related to male fertility, and the PLA2 deficiency in the head of spermatozoa may be one of the reasons causing male infertility. The methods detecting PLA2 content in seminal plasma and the head of spermatozoa can provide powerful evidences for exploring the mechanism of male infertility.