严重急性呼吸综合征康复人员并发股骨头缺血性坏死血清相关蛋白研究

The proteomics research on relational expressed serum proteins among the recovered SARS patients complicating avascular necrosis of femoral head

目的分析严重急性呼吸综合征（SARS）康复人员并发股骨头缺血性坏死血清差异蛋白，探讨其发生机制。方法设立正常健康女性人员为正常对照组（A组，10例）、SARS康复无并发症的女性人员为单纯组（B组，10例）、SARS康复并发股骨头缺血性坏死的女性人员为坏死组（C组，10例）。应用双向凝胶电泳结合基质辅助激光解吸飞行时间串联质谱技术对A、B、C组进行血清比较蛋白质组学研究；酶联免疫吸附测定血清淀粉样P成分在3个组个体血清样本中的表达；统计分析其特异度和敏感度。结果A、B、C组双向电泳凝胶图蛋白质点数分别为632±28、671±55、688±42，匹配率为85％-95％；共找出18个差异蛋白，经质谱鉴定为甲状腺素运载蛋白、丝氨酸蛋白酶抑制因子、α-1抗胰蛋白酶前体、血清淀粉样P成分等10种差异蛋白；与A组和B组比较，C组甲状腺素运载蛋白A链、白蛋白、前白蛋白、补体C4、纤维蛋白原1、载脂蛋白L低表达，而丝氨酸蛋白酶抑制因子、α-1抗胰蛋白酶前体、血清淀粉样P成分高表达；血清淀粉样P成分在A、B、C组个体血清样本中的含量分别为（0．54±0．30）、（0．83±0．39）、（1．21±0．29）ng／ml；与A组和B组比较，C组血清淀粉样P成分高表达（t值分别为2．27、2．98，P〈0．05）；ROC曲线下面积为0．854，特异度77．8％，敏感度85．2％。结论3组间存在差异表达蛋白，血清淀粉样P成分可能成为SARS康复人员并发股骨头缺血性坏死潜在血清标记物之一。

Objective To seek differentially expressed serum proteins in recovered SARS patients complicating avascular necrosis of femoral head （AVNFH）. Methods 2-DE and MALDI-TOF MS were used to study the comparative serum proteomics among female SARS AVNFH group, female SARS non- AVNFH group and female healthy group. ELISA method was used to detect serum amyloid P component in individual serum; specificity and sensitivity of serum amyloid P component were analyzed. Results Average protein points on 2-DE of 3 groups were 632 ± 28,671 ± 55,688 ± 42 respectively, and the matching rate of protein points was ranged from 85% to 95%; eighteen differentially expressed proteins were discovered including transthyretin, serpin peptidase inhibitor, alpha-1-antitrypsin precursor, serum amyloid P components, etc. Compared to healthy group and SARS non-AVNFH group, transthyretin, C4B3, fibrinogen gamma ,apolipoprotein L, apolipoprotein A-Ⅳ precursor, albumin and prealbumin showed lower expression, inversely serpin peptidase inhibitor, alpha-l-antitrypsin precursor and serum amyloid P components showeds higher expression in serum in the SARS AVNFH necrosis group. The serum amyloid P component in 3 groups were0. 54 ± 0. 30 ng/ml,0. 83 ± 0. 39 ng/ml, 1.21 ± 0. 29 ng/ml respectively. The areas under the ROC curve on serum amyloid P component was 0. 854, the specificity was 77.8% and the sensitivity was 85.2%. Conclusion There were differentially expressed serum proteins in three groups. Serum amyloid P components might be one of the potential biomarkers in serum of recovered SARS patients complicating avascular necrosis of femoral head.