应用血清蛋白质组学筛选子痫前期胎盘组织相关抗原  

Screening preeclampsia-associated placenta antigens by serologic proteome analysis

作　　者：王玉萍[1] 牛秀敏[2] 陈亚琼[1] 韩凌霄[1] 胡春秀[1] 杨振华[1] 高秀霞[1] 

机构地区：[1]武警医学院附属医院妇产科,天津300162 [2]天津医科大学总医院妇产科,天津300052

出　　处：《中国实用妇科与产科杂志》2010年第4期277-279,共3页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：武警医学院面上项目(WY200907)

摘　　要：目的筛选子痫前期胎盘组织相关抗原蛋白,为研究子痫前期发病机制提供线索。方法病例来源于2008年1月至10月天津医科大学总医院妇产科分娩的孕妇,用筛选实验对8例重度子痫前期患者胎盘组织采用双向凝胶电泳(2-DW)的方法提取胎盘蛋白,用免疫印迹技术获得免疫印迹图谱,识别差异反应的蛋白质点,根据差异反应的蛋白质点位置在平行胶上找出匹配的蛋白质点,对相应的蛋白质点进行质谱(MALDI-TOF-MS)鉴定和数据库搜索。用验证实验选择子痫前期患者48例,其中轻度子痫前期24例,重度子痫前期24例。对照组为同期产前检查健康孕妇23例,采用ELISA方法检测子痫前期患者血清中纤维蛋白原beta链抗体、热休克蛋白70抗体的表达水平。结果筛选实验子痫前期患者胎盘组织2-DE分离后与患者自身血清免疫印迹反应,而与正常孕妇血清未见免疫印迹反应的蛋白点,经质谱鉴定分别为纤维蛋白原beta链(FBG-β)、热休克蛋白70(HSP70)、线粒体核糖体蛋白的可溶性结构蛋白以及细胞角蛋白9。验证实验证实FBG-βAb、HSP70Ab在子痫前期患者血清中表达升高。结论子痫前期患者体内存在针对胎盘组织相关抗原的体液免疫。这些胎盘抗原蛋白可能在子痫前期的发病中起重要作用。Objective Screening preeclampsia（ PE）-associated placenta antigens in order to provide some clues for the pathomechanism. Methods The samples were pregnancy women,who delivered in the General Hospital of Tianjin Medical University between Jan. 2008 and Dct. 2008. The placenta tissue protein of 8 preeclampsia patient were isolated with techniques of Two-dimensional gel electrophoresis （2-DE）. The immunoblotting imaging film were obtained and the differential reacting protein spots were recognized. Then the differential reacting protein were obtained in the replica gel by matching analysis, and identified by matrix assisted laser desorption ionization time of flight mass spectrometry（ MALDI- TOF-MS） and database searching. Some antibody to antigen proteins among serum were further validated. The serum samples were collected from 48 preelampsia including 24 mild preeclampsia and 24 serious preeclampsia patients, 23 nor- mal pregnant women as control group. The levels of fibrinogen beta chain Ab and Hsp70 Ab were measured using enzyme- linked immunosorbent assay. Results Some proteins from placenta tissue of PE reacted with the sera of PE, but didnt react with the sera of normal. These proteins was identified by MS as fibrinogen beta chain, heat shock protein70, Solution Structure of The Mitochondrial Ribosomal Protein, cytokeratin9. The concentration of fibrinogen beta chain Ab and heat shock protein 70 Ab were higher in the preeclampsia than normal pregnancy. Conclusion The findings indicated that patients with PE develop a humoral response against placenta antigens and these antigens may play an important role for pathogenesis.

关 键 词：子痫前期 双向凝胶电泳 胎盘 免疫 

分 类 号：R71[医药卫生—妇产科学]