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目的比较类风湿关节炎 (rheumatoid arthritis, RA)与骨性关节炎（ osteoarthritis, OA）成纤维样滑膜细胞 (fibroblast like synoviocytes, FLS)蛋白酪氨酸磷酸化状态的差异。方法滑膜细胞原代培养 ,流式细胞仪鉴定滑膜细胞型别 ,提取蛋白进行 SDS－ PAGE单向电泳和 ICE－ PAGE双向电泳分离后，应用抗蛋白酪氨酸磷酸化抗体进行 western blot检测。结果滑膜细胞原代培养至第三代时， CD3、 CD14、 CD20、 CD11b、 von Willibrand factor阳性细胞比例小于 1％ ,RA FLS蛋白酪氨酸磷酸化程度是 OA FLS的 4～ 6倍。结论滑膜细胞原代培养至第三代时即可以获得型别均一的 FLS,RA FLS酪氨酸磷酸化程度较 OA FLS明显增高。
Objective To compare the protein tyrosine phosphorylation status of rheumatoid arthritis (RA) and osteoarthritis (OA) fibroblastlike synoviocytes (FLS). Methods Primary synoviocyte culture was used to identify synovial cell types by flow cytometry. Proteins were extracted for SDS-PAGE and ICE-PAGE two-dimensional electrophoresis. Western blot was performed using anti-protein tyrosine phosphorylation antibody . Results The proportion of CD3, CD14, CD20, CD11b and von Willibrand factor positive cells was less than 1% in primary culture of synovial cells, and the level of tyrosine phosphorylation of RA FLS protein was 4 ~ 6 times higher than that of OA FLS. Conclusions Synovial cells from primary culture to the third generation can obtain homogeneous FLS. The tyrosine phosphorylation of RA FLS is significantly higher than that of OA FLS.