GM-CSF对类风湿性关节炎患者单核细胞合成MMP的影响

Stimulating effect of GM-CSF on the production of matrix metalloproteinases by monocyte/macrophage from patients with rheumatoid arthritis

观察GM-CSF刺激前后,类风湿性关节炎(RA)患者单核/巨噬细胞基质金属蛋白酶(MMP-2、MMP-9)合成及CD147表达的变化。从RA患者外周血及滑液分离单核细胞。RPMI1640培养液中培养,加入不同剂量的GM-CSF,用明胶酶谱法测定刺激24h、48h、72h后MMP-2、MMP-9的活性。用流式细胞术测定刺激前后细胞表面CD147表达量。GM-CSF刺激后,细胞呈多形性的巨噬细胞样改变,MMP-2,MMP-9合成增加,而且这种刺激作用与GM-CSF的剂量相关,100ng/ml时刺激作用显著高于12.5ng/ml及25ng/ml;而与作用时间无显著相关性;滑液单核细胞MMP-2、MMP-9表达水平高于外周血单核细胞。GM-CSF刺激后,单核细胞表面CD147的表达显著高于刺激前;滑液单核细胞表达量高于外周血。GM-CSF刺激后,RA患者外周血及滑液单核细胞MMP-2、MMP-9表达增加,同时伴随细胞表面CD147表达增加。

To observe the changes in the synthesis of the matrix proteinases （MMP-2, MMP-9） and the expression of CD147 by the stimulation with GM-CSF, the monocytes/macrophages were isolated from peripheral blood and synovial fluid of patients with rheumatoid arthritis （RA） by means of the Ficoll-Hypaque technique, and then incubated in RPMI 1640 with different amounts of GM-CSF. The activities and contents of MMP-2 and MMP-9 were evaluated by gelatin-zymography at 24, 48 and 72 hrs respectively, and the expression of CD147 on the surface of monocyte/macrophage was assessed by flow cytometry. It was found that the production of MMP-2 and MMP-9 by monocytes from peripheral blood and synovial fluid of patients with RA was enhanced significantly when stimulated with GM-CSF. This enhancement of production of MMP-2 and MMP-9 correlated with the dose of GM-CSF, but not related with the time of stimulation. Meanwhile, the expression of CD147 on the surface of monocytes of patients with RA was also enhanced by the stimulation with GM-CSF. These results show that the synthesis of MMP-2 and MMP-9 as well as the expression of CD147 were enhanced significantly after stimulation with GM-CSP, indicating a potential role of this phenomenon in the pathogenesis of rheumatoid arthritis.