Mdivi-1对膝骨关节炎患者滑膜成纤维细胞增殖的抑制作用及可能机制

Role of Mdivi-1 in inhibiting proliferation of synovial fibroblasts and its possible mechanism in patients with knee osteoarthritis

目的探讨线粒体分裂抑制剂-1(mitochondrial division inhibitor-1,Mdivi-1)在骨关节炎患者滑膜成纤维细胞增殖中的抑制作用及可能机制。方法取膝骨关节炎患者关节置换术中切除的滑膜组织,提取原代滑膜成纤维细胞进行传代培养,取对数生长期细胞随机分为观察组和对照组,分别应用摩尔浓度为50μmol/L Mdivi-1溶液、体积分数0.1%DMSO溶液干预24 h,采用MitoTracter Green染色法观察2组细胞线粒体形态,CCK-8法检测细胞增殖,DHE染色法检测活性氧表达,实时荧光定量PCR法检测环氧化酶2(cyclooxygenase-2,COX-2)mRNA、基质金属蛋白酶-13(matrix metalloprotein-13,MMP-13)mRNA相对表达量。结果干预24 h,与对照组比较,观察组滑膜成纤维细胞线粒体片段减少,长度明显延长。干预24 h,观察组滑膜成纤维细胞增殖吸光度值(1.13±0.10)、活性氧表达吸光度值(0.037±0.008)、COX-2 mRNA相对表达量(0.69±0.11)、MMP-13 mRNA相对表达量(0.54±0.21)均低于对照组(1.38±0.62、0.070±0.012、1.00±0.00、1.00±0.00)(P<0.05)。结论Mdivi-1可抑制膝骨关节炎滑膜成纤维细胞增殖,其机制可能与抑制活性氧过量产生及COX-2、MMP-13表达有关。

Objective To investigate the role of mitochondrial division inhibitor-1(Mdivi-1)in inhibating the proliferation of synovial fibroblasts in patients with knee osteoarthritis(OA)and its possible mechanism.Methods The primary synovial fibroblasts were extracted from the resected knee synovial tissues in arthroplasty in OA patients for subculture,and the cells in logarithmic growth phase were randomly divided into observation group treated with 50μmol/L Mdivi-1 and control group treated with 0.1%DMSO for 24 h.After 24 h of culture,MitoTracter Green was used to detect the mitochondrial morphology,CCK-8 was used to detect the cell viability,DHE staining was used to detect the reactive oxygen species production,and real-time fluorescence quantitative PCR was used to detect the expressions of cyclooxygenase-2 and matrix metalloprotein-13 mRNAs.Results After 24 h of culture,mitochondria decreased in fragment and extended in length in synovial fibroblasts.The optical densities of cell proliferation and reactive oxygen species,as well as the relative expressions of cyclooxygenase-2 and matrix metalloprotein-13 mRNAs were lower in observation group(1.13±0.10,0.037±0.008,0.69±0.11,0.54±0.21)than those in control group(1.38±0.62,0.070±0.012,1.00±0.00,1.00±0.00)(P<0.05).Conclusion Mdivi-1 can inhibit the proliferation of synovial fibroblasts,probably by inhibiting the reactive oxygen species,cyclooxygenase-2 and matrix metalloprotein-13 in knee OA patients.