人表皮聚角蛋白微丝蛋白的提取及其在类风湿关节炎诊断中的应用

The extraction of filaggrin and its significance in rheumatoid arthritis

目的 建立从人表皮提取聚角蛋白微丝蛋白 (filaggrin)抗原的方法 ,检测抗filaggrin抗体 (AFA)在类风湿关节炎 (RA)中的分布 ,探讨AFA对早期RA诊断的价值。方法 从人表皮细胞中提取并部分纯化filaggrin抗原 ,经SDS PAGE后用免疫印迹法检测 (Westernblot) 10 3例类风湿关节炎 (RA)血清和 140例对照血清 ,包括系统性红斑狼疮 (SLE)、干燥综合征 (SS)、骨关节炎 (OA)等。结果 本研究方法提取的filaggrin能作为AFA免疫印迹检测的抗原。 10 3例RA病人中AFA敏感性和特异性分别为 35 9%、93 7% ,与疾病对照组和正常对照组相比 ,差异有显著性 (P <0 0 5 )。结论 从人表皮提取和部分纯化filaggrin抗原 ,能满足免疫印迹检测的要求。用免疫印迹法检测RA患者的AFA抗体 ,对RA有较好的敏感性和特异性 ,可临床用于RA的诊断。由于基因的变异性 ,不同个体的filaggrin表达强弱不同。

Objective To establish the method for extracting the cytokeratin filament aggregating protein (filaggrin) from human epidermis,to detect the anti filaggrin antibody (AFA) in rheumatoid arthritis (RA) and to determine the diagnostic value of AFA in RA.Methods Filaggrin was extracted from human epidermis and partially purified.AFA in 103 patients with RA and 140 controls was detected by Western blotting.Results Filaggrin extracted from the epidermis could be used as the antigen in AFA test by Western blotting.AFA test resulted in diagnosis of 35 9% of 103 RA samples,with a specificity of 93 7%.The result was significantly higher compared with patients′ and normal controls ( P <0 05).Conclusion Filaggrin extracted from the human epidermis could be used in Western blotting.AFA has diagnostic value in RA.The individuality expresses heterogeneous levels of filaggrin for the gene variation.