应力性骨折的血清蛋白质组学分析

Analysis of serum proteome of patients with stress fracture

目的采用双向电泳-串联飞行质谱技术分析应力性骨折(SF)患者的血清蛋白质组变化,筛选可用于SF早期诊断的生物标志物。方法 SF组和正常对照组血清样本均为混合血清,各由21例平均年龄为19岁的SF战士和正常对照战士的血清分别混合而成。血清样本去除高丰度蛋白后进行双向凝胶电泳(2-DE),比较两组血清蛋白质谱图,寻找差异蛋白点。采用基质辅助激光解吸串联飞行时间质谱(MALDI-TOF/TOF)技术,结合生物学软件和数据库检索对差异蛋白点进行鉴定。结果初步筛选出7个差异蛋白点,经质谱鉴定得到6种蛋白质,分别与新陈代谢、维持正常渗透压、免疫应答及氧化反应相关。在SF组中,对氧磷酶1、白蛋白对碘氧基苯甲醚CRA_h、血清白蛋白前体、白蛋白、甲状腺素结合前白蛋白表达量减少(P<0.05),而血红素结合蛋白前体表达量增加(P<0.05)。结论基于凝胶电泳的蛋白质组学分析可发现与SF相关的血清生物标志物;机体的免疫应答及氧化反应系统可能参与了SF的发生发展。

Objective To analyze the serum proteomic changes in stress fracture （SF） patients by two-dimensional gel electrophoresis （2-DE） and matrix-assisted laser desorption ionization time of flight （MALDITOF） analysis,and to screen potential biomarkers for the early diagnosis of SF.Methods Pooled serum samples were taken from either 21 SF patients （SF group） or 21 normal healthy soldiers （control group）.All the subjects were male soldiers with average age of 19 yr.The serum samples were then analyzed by 2-DE after removal of high abundant albumin and immunoglobulin.The protein finger printings were compared between SF and control group,and the different positive protein spots were identified by MALDITOF combined with bioinformatics software GPS-MASCOT and database NCBInr.Results Seven different protein spots were primarily screened out,of which six proteins,correlated with metabolism,osmoregulation,immune response and oxidation,were identified by MALDITOF analysis.Compared with those of the control group,the expression of five proteins,including paraoxonase 1,albumin isoform CRA_h,serum albumin precursor,albumin and thyroxine binding prealbumin,were down-regulated （P〈0.05）,and hemopexin precursor was up-regulated in SF group （P〈0.05）.Conclusion The gel electrophoresis-based proteomic analysis may be helpful to discover the potential serum biomarkers of SF,and the immune response and oxidation reaction may participate in the occurrence and development of SF.