燃煤污染型砷中毒肝损伤患者血清差异蛋白质的筛选

目的 对比分析燃煤污染型砷中毒肝损伤患者和健康人血清中差异表达的蛋白,筛选与燃煤污染型砷中毒致肝损伤发生密切相关的血清蛋白质.方法 6例血清标本来自于贵州省兴仁县交乐病区燃煤型砷中毒肝损伤患者及病区健康人(对照组),采用双向凝胶电泳(2-DE)分离血清中总蛋白,硝酸银染色后经图像分析识别差异表达的蛋白质,基质辅助激光解吸电离飞行时问质谱(MALDI-TOF-MS)鉴定差异表达的蛋白质点,寻找与燃煤污染型砷中毒致肝损伤有关的蛋白质.结果成功地建立了血清2-DE图谱.差异蛋白分析显示,砷中毒肝损伤组平均蛋白点数为(824±31)个,对照组为(782±42)个,两组血清2-DE图谱的匹配率为94.9%(782/824).筛选出两组间的差异蛋白点49个,对其中表达量差异3倍以上的30个蛋白点进行MALDI-TOF-MS分析,鉴定出相关蛋白10个.与对照组比较,其中α2-巨球蛋白、B细胞受体相关蛋白31、细胞角蛋白1、载脂蛋白A-I在砷中毒肝损伤组表达上调,触珠蛋白、α2-HS-糖蛋白、细胞外信号调节激酶4、锌指蛋白323、ZAP-70、SP40表达下调.结论成功地建立了分辨率高、重复性较好的燃煤污染型砷中毒肝损伤患者血清2-DE图谱,并筛选出一些与健康人血清中差异表达的蛋白质,这些蛋白质能否作为燃煤污染型砷中毒肝损伤诊断的血清标志物还有待于进一步验证.

Screening of differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.