以FSH受体的配体结合部位作为抗原进行主动免疫对大鼠生殖的影响  

Effect on Fertility by Active Immunization with Antigen Targgeting FSH Receptor Ligand Binding Region in Rats

作　　者：石燕[1] 郁琳[1] 朱燕[1] 蒋雅红[1] 孙兆贵[1] 王健[1] 

机构地区：[1]上海市计划生育科学研究所,国家人口和计划生育委员会计划生育药具重点实验室,上海200032

出　　处：《实验动物与比较医学》2013年第1期12-18,共7页Laboratory Animal and Comparative Medicine

基　　金：上海市科学技术委员会项目（10140901502）资助

摘　　要：目的重组表达FSHR抗原主动免疫SD大鼠，探讨FSHR免疫抑制对大鼠卵巢功能的影响，以构建免疫异常导致的卵巢早衰（POF）大鼠模型。方法以FSHR分子N端配体受体结合区片段为靶位，制备重组抗原，同时针对免疫原性预测位点人工合成3段肽，以佐剂为对照，免疫大鼠，免疫后检测血清中的FSHR抗体滴度。在体外利用免疫荧光技术检测抗血清对卵丘复合体中FSHR的识别能力：并且研究FSHR主动免疫后卵巢对PMSG反应性的变化；检测血清FSH和雌激素水平，并与雄鼠交配，观察FSHR主动免疫对窝仔数的影响。结果主动免疫后血液中产生高滴度水平的抗体，在免疫周期后持续6周无明显下降。主动免疫产生的抗体可以与颗粒细胞特异性结合：同时，卵巢对促性腺激素的反应性降低。重组蛋白免疫组的大鼠动情周期紊乱，发情期缩短，血液FSH水平上升，而雌激素水平降低，与正常雄性交配窝仔数下降。结论人工制备大鼠的FSHR免疫原，主动免疫大鼠，导致大鼠血液出现高水平的FSHR抗体，可以与卵巢颗粒细胞上的FSHR特异结合，导致其对促性腺激素的反应性降低，进而引发大鼠出现卵巢早衰的症状。Objective To establish the rat model of premature ovarian failure by immune suppression to FSH receptor （FSHR）, and to evaluate their phenotype. Methods On the basis of biological informa- tion of the whole FSHR protein structure, the extracellular domain of FSHR was cloned. The recombi- nant protein obtained by prokaryotic expression and three synthetic peptides of the protein were used to immunize 4 weeks old female SD rats. Estrous cycle, and blood FSH, estrogen and progesterone were detected separectely. Results After the active immunization, high titer antibodies were detected in the blood and lasted for 6 weeks. The antibodies were shown to specifically bind with the granulosa cells in vitro and at the same time, the ovarian response to gonadotropins was reduced in vivo. Estrous cycle disorders with shortened estrus, blood FSH rise and estrogen decrease were detected in rats immunized with recombinant protein; and the litter size was decreased while mating with normal males. Conclusion The active immunization in rats against FSHR ligand binding sites resulted in high titers of FSHR antibodies in the blood, which could specifically bind to ovarian granulosa cells, and resulted in lower response to gonadotropin in ovaries, and then symptoms of premature ovarian failure were observed in rats.

关 键 词：FSHR 主动免疫 卵巢早衰模型 SD大鼠 

分 类 号：R711.75[医药卫生—妇产科学] R-332[医药卫生—临床医学]