精浆蛋白质电泳两种分析方法的探讨

目的:探讨琼脂糖电泳和SDS-琼脂糖电泳分析精浆蛋白质的临床可行性。方法:69例不育患者精液标本按常规和特殊检查结果分为弱畸精子症(n=19)、弱精子症(n=22)和相对正常(n=28)3组,然后离心分离获取精浆。分别采用琼脂糖和SDS-琼脂糖作介质,不同的点样时间、迁移功率,酸性结晶紫和氨基黑两种染色,对精浆蛋白质进行检测,将电泳图谱进行吸光度仪扫描并将结果对比分析。结果:以SDS-琼脂糖作介质,酸性结晶紫染色可将精浆分为4条带,但区带弥散,相互间拖尾严重;以琼脂糖作介质,pH 9.2碱性缓冲条件,点样6 m in,可将精浆分为A、B、C、D、E、F 6条带,氨基黑染色后,区带清晰,间隔适当,易于吸光度仪分析相对含量,重复性好。电泳结果经方差分析,弱精子症组与相对正常组、弱畸精子症组在C与E带上均有差别,而相对正常组与弱畸精子组无差别;同时,相对正常组标本之间的电泳图谱也有显著的不同。结论:以琼脂糖作介质,在碱性缓冲条件下电泳,氨基黑染色来分离检测精浆蛋白质具有较高的可行性,该方法操作简单、快速,可适合临床常规开展。

Study of Two Electrophoresis Procedures of Seminal Plasma Proteins

Objective: To analyse clinical feasibility of two electrophoresis procedures of seminal plasma proteins,agarose gel electrophoresis and SDS-agarose gel electrophoresis.Methods: Sixty-nine semen samples were examined and classified into three groups: the asthenozoospermia(n=22),the asthenoteratozoospermia(n=19),and the relative normal group(n=28) with normal routine and special test results,according to WHO routine and special test criterion.Then,the seminal plasma protenis were separated by two different electrophoresis,with SDS-agarose and agarose support medium,the buffer pH(7.0) and(9.2) respectively.The agarose gel electrophoresis was done under various sample loading time,motion power and staining modules.The completed gels were scanned and compared the each other statistically.Results: Seminal plasma proteins can be separated into 4 strips by SDS-agarose gel electrophoresis with acid crystal violet,and the strips were diffusion and with dark background.However,6 clear strips named A, B,C,D,E,and F can be obtained by agarose gel electrophoresis with 6 min.After samples were loaded and stained by amidoblack,there showed appropriate spaces among strips,and it was very easy to scan the drying gel by a densitometer.Using agarose gel electrophoresis,the statistical difference in strip C and E was shown between the asthenozoospermia and the relative normal group,and between the asthenozoospermia and the asthenoteratozoospermia,however,not between the relative normal and the asthenoteratozoospermia group.Moreover,the samples in the relative normal group with normal routine and special test results were in different electrophoresis maps.Conclusion: Agarose gel electrophoresis of seminal plasma proteins with buffer pH 9.2,6 min.sample loading and amidoblack stain was a simple,fast and fit technique for clinic.