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目的 从人精浆中分离提取精浆免疫抑制因子 (SPIF)。方法 取常规检测正常精液 5 0份 ,离心分离精子和精浆 ,将精浆低温高速离心 ,取上清液 ,经SephadexG - 10 0层析 ,收集第 1峰 ,作DEAE纤维素离子交换层析 ,收集第 2峰 ,浓缩 ,经SDS -PAGE分析呈单带。在体外对其生物活性进行了测定 ,并以其为抗原经ELISA间接法对 10 0例不育男子和 5 0例生育男子精浆中抗SPIFIgG和IgA进行了测定。 结果 测定分子量为 5 2kd。当浓度分别为 5 0、10 0、2 0 0 μg/ml时 ,对PHA诱导的人外周血淋巴细胞转化抑制率、NK细胞活性抑制率分别为2 7.1%、33.9%、6 6 .9%和 2 3.7%、32 .4%、5 7.4%。ELISA结果 ,不育组与生育组间抗SPIFIgG和IgA分别为38%、2 6 %和 6 %、2 %。结论 该实验提取的 5 2kd的蛋白达到电泳纯且在体外具有一定的生物活性。ELISA间接法结果显示 ,不育组与生育组间存在显著性差异 (P <0 .0 5 )。
Objective To isolate and extract seminal plasma immunosuppressive factor (SPIF) from human seminal plasma. Methods Fifty normal sperm samples were collected and centrifuged to separate sperm and seminal plasma. The seminal plasma was centrifuged at high speed and centrifuged. The supernatant was collected and the first peak was collected by Sephadex G - 10 0 chromatography. DEAE cellulose ion exchange chromatography , The second peak was collected, concentrated and analyzed by SDS-PAGE for a single band. Its biological activity was tested in vitro and anti-SPIFIgG and IgA in seminal plasma of 100 infertile men and 50 male fertilized men were measured by ELISA indirect method. The result of determination of molecular weight of 5 2kd. When the concentrations were 50, 100 and 200 μg / ml respectively, the inhibitory rates of PHA-induced human peripheral blood lymphocyte transformation and NK cell activity were 21.1%, 33.9% and 66.9%, respectively And 2 3.7%, 32.4%, 5 7.4%. The result of ELISA showed that the anti-SPIFIgG and IgA were 38%, 26% and 6%, 2% respectively in infertility group and fertility group. Conclusion The 5 2 kd protein extracted from this experiment reached electrophoretic purity and had certain biological activity in vitro. ELISA indirect results showed that there was a significant difference between sterile group and fertility group (P <0.05).