人膝骨关节炎血清生物学标志物的筛选

目的: 比较膝骨关节炎患者与正常人血清蛋白质组的差异,筛选其能用于骨关节炎诊断、治疗或发病机制研究的血清生物学标志物。方法: 利用双向荧光差异凝胶电泳技术比较膝骨关节炎患者血清(4例)与健康志愿者血清(4例)蛋白图谱的差异,使用基质辅助激光解吸电离-飞行时间质谱及生物信息学相关技术对获得的差异蛋白进行鉴定和初步分析,并用Western blotting进一步验证所鉴定的蛋白。结果: 成功建立了骨关节炎血清差异蛋白质组学的研究方法;通过质谱分析初步鉴定出8个差异蛋白,其中5个蛋白在膝骨关节炎组表达上调,3个蛋白在膝骨关节炎组表达下调。Western blotting进一步验证得到α₂-巨球蛋白在膝骨关节炎组表达上调,与双向荧光胶内差异凝胶电泳技术联合质谱分析的结果一致。结论: 膝骨关节炎患者血清与正常人血清蛋白表达存在明显差异,其中α₂-巨球蛋白可作为骨关节炎潜在的疾病相关生物学标志物进一步研究,为骨关节炎的诊断、治疗和发病机制的研究提供新的线索和实验基础。

Analysis of serum biomarkers in human knee osteoarthritis

AIM: To analyze the proteomic components of the sera from knee osteoarthritis patients and normal people,and to search proteins that might serve as serum biomarkers for osteoarthritis diagnosis,treatment or pathogenesis.METHODS: Two-dimensional fluorescence difference gel electrophoresis(2D-DIGE) was applied to analyze the sera obtained from the patients with knee osteoarthritis(n=4) and normal controls(n=4).The differentially expressed proteins were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS).Western blotting analysis was applied to confirm the results.RESULTS: Comparative proteomic data of serum from the patients with osteoarthritis was successfully obtained.Eight differentially expressed protein spots were observed.Five up-regulated and 3 down-regulated proteins were identified.Western blot analysis confirmed that α2-macroglobulin was increased.CONCLUSION: There are significant differences between serum proteins obtained from the patients with knee osteoarthritis and normal controls.α2-Macroglobulin might be utilized as potential biomarkers for the diagnosis and treatment of osteoarthritis.