AIM: To analyze the proleomic componenls of the sera from knee osleoarthrilis patients and normal people, and to search proleins that might serve as serum biomarkers for osleoarlhrilis diagnosis, trealmenl or palhogenesis. METHODS: Two -dimensional fluorescence difference gel eleclrophoresis (2D - DIGE) was applied lo analyze the sera oblained from the palienls with knee osleoarlhrilis (n =4) and normal controls (n =4). The differentially expressed proleins were idenlified by malrix - assisted laser desorplion ionizalion lime - of - flighl mass speclromelry ( MALDI - TOF -MS). Western blotting analysis was applied lo confirm the resulls. RESULTS; Comparalive proleomic data of serum from the palienls wilh osteoarlhrilis was successfully oblained. Eight differentially expressed prolein spots were observed. Five up - regulaled and 3 down - regulated proleins were identified. Weslern blot analysis confirmed that α2 - macroglobulin was increased. CONCLUSION: There are significant differences between serum proleins obtained from the patienls with knee osleoarlhrilis and normal controls. Α2 - Macroglobulin might be utilized as potenlial biomarkers for the diagnosis and -realmenl of osteoarlhrilis.%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.
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