老年性骨关节炎与类风湿关节炎滑膜细胞转化特性的比较研究

Comparative study of transformation characteristics of synoviocytes in senile rheumatoid arthritis and osteoarthritis

目的与骨关节炎比较,探讨类风湿关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)胞外信号活化蛋白激酶(ERK)的活化状态和P53蛋白的异常表达。方法原代培养RA FLS和OA FLS,应用western blot检测二者ERK活化状态的差异;应用抗P53抗体(DO-1和PAb240)与抗PCNA抗体对RA和OA的滑膜组织和细胞进行免疫组织化学染色;并进一步应用western blots确定RA FLS P53的异常表达。结果与OA相比,RA滑膜衬里层P53蛋白表达增多,主要为与抗突变型P53抗体阳性反应细胞积聚,并进一步证实RA衬里层成纤维样滑膜细胞P53表达异常;而且RA FLS在低血清和非贴壁情况下ERK活化状态均明显高于骨关节炎。结论类风湿关节炎衬里层成纤维样滑膜细胞P53蛋白异常表达,而且ERK处于持续活化状态,这为RAFLS的转化特性提供了新的佐证。

Objective To investigate the activation status of extracellular signal regulated protein kinase （ERK） and abnorma] expression of P53 protein in fibroblast-like synoviocytes （FLS） in rheumatoid arthritis （RA）, by comparing with those studies in osteoarthritis （OA）. Methods RA FLS and OA FLS were primarily cultured. The activation status of ERK in different cells was detected using Western blotting. Immunohistochemical staining in both RA and OA synovia] membrane and synoviocytes was performed using antibodies against P53 （ DO-1, PAb240） and anti-PCNA. The abnormal expression of P53 in RA FLS was detected using Western blotting. Results The expression of P53 protein in the synovial lining layer in RA was much more than that in OA, which was mainly the accumulation of reacting cells with anti-mutant phenotype P53 antibody positive. The abnormal expression of P53 protein in RA FLS lining layer was further confirmed. The activation status of ERK in RA FLS in the conditions of low-serum and non-adherence was much higher than that in OA FLS. Conclusion The expression of P53 protein in RA FLS lining layer is abnormal, and activation status of ERK is continuous. This provides new evidence for the transformation characteristics of RA FLS.