结缔组织生长因子对类风湿关节炎成纤维样滑膜细胞增殖的影响

Effect of Connective Tissue Growth Factor on Proliferation of Fibroblast-like Synoviocytes of Rheumatoid Arthritis

目的观察结缔组织生长因子(CTGF)对体外培养的类风湿关节炎成纤维样滑膜细胞(FLS)增殖的影响,探索CTGF在类风湿关节炎滑膜病变中的可能作用机制。方法体外培养FLS并用免疫组化法鉴定。加入不同浓度的外源性CTGF[0 ng/m L(对照组),5,10,25,50 ng/m L],用3H-Td R掺入法检测对FLS增殖的影响。特异性细胞信号通路阻断剂预处理后,探讨CTGF对FLS作用的可能机制。结果体外培养FLS,经光镜观察及免疫组化鉴定符合成纤维样滑膜细胞特征。CTGF 10,25,50 ng/m L组细胞增殖随CTGF浓度上升而增加,且与对照组比较差异具有统计学意义(P<0.01);但CTGF 25 ng/m L组与50 ng/m L组间差异无统计学意义(P>0.05)。特异性的信号通路阻断剂分别阻断NF-κB信号通路、PI-3激酶信号通路、p38 MAPK信号通路和ERK-1/2信号通路,只有PD98059组FLS增殖较CTGF 25 ng/m L组有明显差异(P<0.05)。结论外源性CTGF可以促进体外培养的FLS增殖,并且在一定浓度范围内呈现浓度依赖性。这种促进增殖的作用很可能是通过活化ERK-1/2信号通路完成的。

Objective To investigate the in vitro effect of connective tissue growth factor （CTGF） on the proliferation of fibroblast-like synoviocytes （FLS） of rheumatoid arthritis （RA）, and explore the potential signal transduction mechanisms of CTGF-induced FLS proliferation. Methods FLS were cultured in vitro and identified by immunohistochemieal method. FLS proliferation was measured by thymidine incorporation after exogenous CTGF stimulation with different concentrations （0,5, 10,25,50 ng/mL）. After specific cellular signaling pathway blockers pretreatment of cultured FLS, observe the effects of CTGF on FLS proliferation by thymidine incorporation. Results Cells cultured in vitro were complied with the FLS characteristics by optical microscopy and identification. The proliferation of FLS was gradually increased as the concentration of CTGF elevating from 10 ng/mL to 50 ng/mL, which were significantly higher than the control group （P 〈 0.01 ）. However, no significant difference was observed between FLS cultured with 25 ng/mL CTGF and those cultured with 50 ng/mL CTGF （P 〉 0.05 ）. Inhibition of ERK- 1/2 by PD98059 significantly suppressed CTGF-mediated FLS proliferation （P 〈 0.05 ）. Conclusion CTGF induced a proliferative response in FLS, and this action is likely dependent on the activation of ERK-1/2 signal pathway.