论文部分内容阅读
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 To screen placenta tissue related antigen protein in preeclampsia and provide clues for studying the pathogenesis of preeclampsia. Methods From January 2008 to October 2008, Tianjin Medical University General Hospital Obstetrics and Gynecology, pregnant women, with screening experiments in 8 cases of severe preeclampsia placental tissue using two-dimensional gel electrophoresis (2-DW) extraction of placenta Western blotting was used to obtain the immunoblot. The differentially reacted protein spots were identified. The matched protein spots were found on the parallel gels according to the differential reaction spots. The corresponding protein spots were identified by mass spectrometry (MALDI-TOF-MS) Identification and database search. Forty-eight patients with preeclampsia were selected by validation experiment, including 24 mild preeclampsia and 24 severe preeclampsia. In the control group, 23 pregnant women were examined prenatally. The serum levels of fibrinogen beta chain and HSP70 in preeclampsia were detected by ELISA. Results The placental tissues from preeclampsia patients were separated by 2-DE, and their serum immunoblots were detected by Western blotting. The spots that were not immunoblotted with the serum of normal pregnant women were identified by mass spectrometry as fibrinogen beta chain (FBG-β ), Heat shock protein 70 (HSP70), soluble structural protein of mitochondrial ribosomal protein, and cytokeratin 9. Validation experiments confirmed that FBG-βAb, HSP70Ab expression in preeclampsia patients increased. Conclusion There is humoral immunity against placenta tissue associated antigen in preeclampsia patients. These placental antigen proteins may play an important role in the pathogenesis of preeclampsia.