TNF-α上调单核巨噬细胞MMP-9的活性与类风湿关节炎关节破坏的关系

Association of TNF-α upregulation of MMP-9 activation in monocyte-derived macrophages with progression of joint damage in patients with rheumatoid arthritis

目的:探讨TNF-α对单核巨噬细胞基质金属蛋白酶9(MMP-9)的表达与酶活性的影响以及与类风湿关节炎患者关节破坏的关系。方法:用双抗体夹心ELISA法检测类风湿关节炎患者组(RA)和对照组血清和关节滑液中TNF-α、MMP-9的含量,观察MMP-9与X线表现积分(Larsen)的关系。体外将佛波酯(TPA)和不同浓度(0、1、10、20μg/L)TNF-α共同孵育THP-1细胞24 h后,运用Western blotting方法检测MMP-9蛋白的表达,明胶酶谱法检测MMP-9活性,侵蚀小室法观察分化前后THP-1细胞的侵蚀力。结果:RA患者组血清和关节滑液中TNF-α、MMP-9的水平明显高于对照组(P<0.05),且血清和滑液MMP-9与Larsen积分显著相关(r=0.37和r=0.32,P<0.01);体外细胞实验中,TNF-α上调分化的THP-1中MMP-9的表达和酶活性,并且增强分化的THP-1细胞的侵蚀性,并与TNF-α呈浓度依赖性。结论:TNF-α上调单核巨噬细胞MMP-9表达及活化,增强了炎症细胞的侵蚀力,可能在RA关节破坏机制中起着重要的作用。

AIM： To explore the expression and activation of matrix metalloproteinase- 9 （MMP- 9） from monocyte - derived macrophages induced by tumor necrosis factor - α （ TNF - α and to investigate its association with progression of joint damage in patients with rheumatoid arthritis. METHODS ： TNF - α and MMP - 9 in serum and synovial fluid from patients with early RA and controls were tested with a double - antibody enzyme - linked immunosorbent assay. The correlation between MMP-9 and Larsen score over the first 12 months was analyzed. THP- 1 cells differentiated by the treatment with TPA were stimulated with increasing concentration of TNF - α for 24 h in vitro. The protein expression of MMP -9 was determined by Western blotting. The activity of MMP -9 was measured by gelatinolytic zymography. Boyden chamber - matrigel in vitro invasion assay was used to detect the invasive capacity. RESULTS： The levels of TNF - α and MMP - 9 in serum and synovial fluid of RA patients were significantly higher than those in controls （ P 〈 0. 05 ）. Serum and synovial fluid levels of MMP - 9 correlated significantly with Larsen score （ r = 0. 37 and 0. 32, P 〈 0.01 ）. The MMP - 9 activity and invasive ability of co - cultured THP - 1 cells with TNF - a and TPA were higher than those of non - TNF -α treatment. CONCLUSION： TNF -αupregulates MMP- 9 activation and promotes infiltration of monocyte -derived mac- rophages, indicating that TNF - α play an important role in the pathogenesis of RA.