双向电泳-质谱技术筛选克山病血清差异蛋白

Screening of differentially expressed proteins in serum from subjects with Keshan disease by two-dimensional electrophoresis and mass and mass spectrometry

目的比较克山病患者与健康者血清蛋白表达谱,探讨克山病发病机制,寻找与克山病发生相关差异蛋白。方法从克山病病区选择8例慢型克山病,病区对照以及非病区对照各8例,应用双向凝胶电泳技术分离克山病患者同健康者的差异蛋白点,基质辅助激光解吸电离-飞行时间质谱鉴定差异蛋白。结果凝胶图像分析显示9个差异蛋白质点,经质谱鉴定确定8种蛋白。3种蛋白在克山病组较非病区健康组上调,其功能主要与脂类代谢,免疫调节,凋亡抑制有关。3种蛋白下调,主要与细胞内铁离子平衡密切相关。克山病组较病区健康组2种蛋白表达上调,主要与蛋白酶抑制等功能有关。结论该研究发现触珠蛋白、血清白蛋白和α1-抗胰蛋白酶、转铁蛋白、α2-HS糖蛋白可作为克山病的诊断与预后的候选生物学指标,具有重要意义。

Objective To screen differentially expressed proteins in serum in patients with Keshan disease（KD）,peripheral blood protein expression spectrum between subjects with Keshan disease and health controls were compared.Methods Differentially expressed protein spots were screened by two-dimensional gel electrophoresis（2-DE） between Keshan disease and health control subjects,and constitutive protein were identified by matrix assisted laser adsorption/ionization-time of flight mass spectrometry（MALDI-TOF-MS）.Results 9 differentially expressed protein spots were showed in 2-DE images and 8 differentially expressed proteins were identified by MALDI-TOF-MS.In them,3 up-regulated proteins,mainly related to lipid metabolism,apoptosis resistance,immunological regulation and 3 down-regulated proteins,involved to cellular iron ion homeostasis;2 up-regulated proteins in serum in patients with KD versus controls from KD areas were detected,mainly associated with protease inhibition.Conclusions Haptoglobin,serum albumin,alpha-1-antitrypsin,transferring and α2-heremans schmid glycoprotein may be considered as candidate biological indicators used for diagnosis or prognosis of KD.