ITRAQ技术筛选系统性红斑狼疮合并股骨头坏死的血清差异蛋白质组学变化

Screening differentially expressed proteins in the serum of patients with systemic lupus erythematosus combined with osteonecrosis of the femoral head by iTRAQ technology

背景:系统性红斑狼疮合并股骨头坏死是由于长期大剂量应用皮质类固醇糖皮质激素造成股骨头活性成分死亡的病理过程,但其早期诊治缺乏特异的生物标志物。目的:应用同位素相对标记与绝对定量技术(i TRAQ)筛选系统性红斑狼疮合并激素性股骨头坏死的血清标志物。方法:分别收集系统性红斑狼疮合并激素性股骨头坏死患者血清8例(实验组)、系统性红斑狼疮使用激素后无股骨头坏死的患者血清(对照组)10例,采用i TRAQ联合二维液相色谱-串联质谱技术(2D-LC-MS/MS)对差异蛋白质进行筛选与鉴定。结果与结论:①共鉴定出10种差异蛋白质,与对照组相比有统计学差异,其中实验组中铜篮蛋白、免疫球蛋白Ig G、免疫球蛋白κ链、补体C2等4种蛋白表达上调;内皮细胞蛋白C受体、人源性激肽释放酶结合蛋白、色素上皮衍生因子、四连接素、胰岛素样生长因子2、甲状腺素运载蛋白等6种蛋白表达下调;②KEGG通路分析提示以上10个蛋白与Wnt信号通路、补体与凝血级联反应、系统性红斑狼疮这3条信号通路密切相关;③结果说明,通过蛋白质组学技术成功得到了可靠的激素性股骨头坏死血清差异蛋白质,为激素性股骨头坏死的诊断及其发病机制提供了理论依据。

BACKGROUND:Systemic lupus erythematosus(SLE)combined with osteonecrosis of the femoral head(ONFH)is a result of long-term application of steroids that induces death of active ingredients in femoral head.But there is a lack of specific biomarker for early diagnosis.OBJECTIVE:To screen the systemic markers of SLE with steroid-induced ONFH using isobaric tags for relative and absolute quantitation(iTRAQ)technology.METHODS:Serum of eight patients with SLE and steroid-induced ONFH was collected and used as the experimental group,and serum of 10 patients with SLE but without steroid-induced ONFH served as the control group.The differentially expressed proteins were screened and identified by iTRAQ technique and two-dimensional liquid chromatography-tandem mass spectrometry.RESULTS AND CONCLUSION:Totally 10 kinds of differentially expressed proteins were identified.There was significant difference between two groups.In the experimental group,the expression levels of copper basket protein,immunoglobulin IgG,immunoglobulinκchain and complement C2 were significantly up-regulated.The expression levels of endothelial cell protein C receptor,human kallikrein-binding protein,pigment epithelium-derived factor,tetracycline,insulin-like growth factor 2,and thyroxine carrier protein were down-regulated.KEEG pathway analysis indicated that 10 kinds of proteins were closely related to Wnt signaling pathway,coagulation and complement cascades reaction signaling pathway and SLE signaling pathway.In summary,the differentially expressed proteins are obtained successfully in steroid-induced ONFH by proteomics,which provides theoretical basis for the diagnosis and pathogenesis of steroid-induced ONFH.