重组Furin蛋白对类风湿关节炎滑膜细胞生物学行为的影响

Effect of recombinant furin protein on biological behavior of synoviocytes in rheumatoid arthritis

[目的]探讨重组弗林蛋白(Furin)对于类风湿性关节炎(rheumatoid arthritis,RA)滑膜细胞增殖、迁移、侵袭和细胞因子分泌的影响。[方法]从类风湿关节炎患者关节内提取RA滑膜组织后培养原代滑膜成纤维细胞(fibroblast-like synovial cells,FLS);将重组Furin蛋白按照不同浓度(250 ng/ml,500 ng/ml)加入RA滑膜细胞培养基中诱导培养并采用MTT、细胞划痕、Transwell、ELISA等方法检测重组蛋白处理对细胞增殖、迁移、侵袭和炎性因子分泌等生物学特性的影响。[结果]与空白对照组比,加入重组Furin蛋白处理24 h、48 h,对类风湿滑膜细胞增殖活动没有明显影响(P>0.05)。处理24 h后与空白对照组比,迁入划痕伤口内细胞数无显著区别(P>0.05),滑膜细胞穿透基底膜细胞数明显减少(P<0.05),且与剂量浓度呈正相关。培养液上清中IL-1α和IL-17含量升高(P<0.05),而各组间IL-1β和TNF-α含量差异无统计学意义(P>0.05)。[结论]重组Furin蛋白诱导可抑制类风湿关节炎滑膜细胞侵袭能力同时可促进其分泌IL-1α和IL-17。

[Objective] To assess the effect of recombinant furin protein on proliferatian, migration, invasion and secretion of synoviocytes in rheumatoid arthritis （RA） . [Method] Fibroblast-like synovial cells were isolated and cultured from synovial tissues harvested from patients with RA during the knee joint replacement. Different concentrations of recombinant furin protein were applied to the culture medium for RA synoviocytes in order to testify effect on proliferation, migration, invasweness and secretion of the synoviocytes by MTT. scratch assay, transwell and enzyme-linked immunosorbent assay for IL-1α, IL-1β, IL-17, as well as TNF-α in the supernatant. [Result] Compared with the control group, proliferation of rheumatoid synovial cells was not affectext for 24h and for 48h （P〉0.05） . After adding recombinant Furin protein, there was no significant difference in number of ceils in the wound （P〉0.05） . However, the number of penetrating synoviocytes decreased significantly （P〈0.05）, associated with positive correlation to the concentration of Furin protein. The optical density value of IL-1α and IL-17 were respectively higher than that of control group （P〈0.05）, while the differences in IL- 1β and TNF-α were not statistically significant （P〉0.05） . [Conclusion] Recombinant furin protein inhibits the synoviocytes＇ invasion, simultaneously improve rheumatoid arthritis synoviocyte to secret IL-1α and IL-17 as well.