精子蛋白32(sp32)/OY-TES-1在生精细胞中的分布及其抗体的血清学检测  

Distribution of Sperm Protein 32(sp32)/OY-TES-1 in Spermatogenic Cell and Its Antibody Sera Survey

作　　者：范蓉[1] 黄巍[2] 肖绍文[3] 罗彬[4] 李颀[4] 徐丽湘[4] 罗国容[4] 谢小薰[4] 

机构地区：[1]广西中医学院组胚教研室,南宁530001 [2]广西中医学院一附院妇产科,南宁530023 [3]广西医科大学一附院神经外科,南宁530021 [4]广西医科大学组胚教研室,南宁530021

出　　处：《生殖与避孕》2009年第6期365-369,共5页Reproduction and Contraception

基　　金：国家自然科学基金(30360026;30760055);广西自然科学基金(0728108;0728148);广西研究生科研创新项目(200810598100M207);广西大型仪器协作共用网测试补助(4982007077;4992007078;6662008079)资助项目

摘　　要：目的:了解精子蛋白(sperm protein32,sp32)/OY-TES-1蛋白在生精细胞中的分布及抗sp32/OY-TES-1抗体在不育症患者血清中的出现情况。方法:用重组表达的OY-TES-1麦芽糖融合蛋白免疫新西兰白兔,制备sp32/OY-TES-1多克隆抗体,且经纯化鉴定后用于检测人精子和大鼠正常睾丸组织;用ELISA试验,检测不育症患者和正常人血清中抗sp32/OY-TES-1抗体。结果:①免疫后的兔血清含高效价的sp32/OY-TES-1多克隆抗体,并可与OY-TES-1麦芽糖融合蛋白特异性地结合;②免疫组化染色显示人精子头部、大鼠精子和精子细胞中呈免疫反应阳性。③ELISA检测结果显示76例正常人血清中无抗sp32/OY-TES-1抗体,而不育症患者血清其抗体的阳性率高达70.77%(46/65),并有较高的抗体水平。结论:本实验制备的sp32/OY-TES-1多克隆抗体可用于大鼠睾丸生殖细胞的检测,为后续用大鼠进行sp32/OY-TES-1的相关研究奠定了基础。sp32/OY-TES-1特异性地表达于精子细胞和精子,其抗体的出现意义与不育症的关系有待进一步研究。Objective： To understand the distribution of sp32/OY-TES-1 in spermatogenic cell and occurrence of its antibody in sera of sterility patients. Methods： New Zealand rabbit was immuned with recombinant fusion protein of maltose binding protein and OY-TES-1 （MBP/OY-TES-1） for preparation of polyclonal antibody of sp32/OY-TES-1. The antibody was purified and identified, and then used for the immunohistochemistry staining to test human sperm and the rat normal testis. The level of the anti-sp32/-TES-1 antibody of sera from the health individuals and patients with infertility was tested by ELISA, respectively. Results： 1） After immunization of rabbit, high titer of antiserum was obtained. The antiserum was proved to be specifically to the sp32/OY-TES-1 because it can be combined to the MBP/OY-TES-1.2） The immunohistochemistry staining with sp32/OY-TES-1 polyantibody showed that there was positive reaction in the head of human spermatozoa as well as spermatozoa and spermatid of rat. 3） The result of ELISA showed that anti-sp32/OY-TES-1 antibody was detected in 46 of 65 （70.77%） sera from patients with infertility. In some of cases anti-sp32/OY-TES- 1 antibody presented higher titer level. While there was no anti-sp32/OY-TES-1 antibody tested in the sera of 76 normal individuals. Conclusion： The anti-sp32/OY-TES- 1 antibody prepared in this study can be used to detect the germ cell line in the rat testis, which provides the basis for further explorenation of sp32/OY-TES-1 in rat. sp32/OY-TES-1 expressed specifically in the spermatid and spermatozoa. The significance of occurrence of anti-sp32/OY-TES-1 antibody and its relationship with infertility need further study.

关 键 词：精子蛋白32 不育症 血清 

分 类 号：R698.2[医药卫生—泌尿科学]