白藜芦醇通过下调MnSOD诱导类风湿性关节炎成纤维样滑膜细胞凋亡

目的探讨锰超氧化物歧化酶(MnSOD)蛋白表达改变在白藜芦醇(Res)介导的类风湿性关节炎成纤维样滑膜细胞(RA-FLSs)的增殖和凋亡中的作用。方法 Res处理RA-FLSs后,CCK-8法检测细胞增殖,Western blot检测MnSOD蛋白表达水平;慢病毒感染RA-FLSs后,8 mg·L^-1的嘌呤霉素筛选感染后细胞,以此建立3种稳定细胞系:MnSOD过表达、MnSOD干扰、MnSOD对照;激光共聚焦显微镜检测每种细胞系在5μmol·L^-1过氧化氢(H_2O_2)和200μmol·L^-1 Res处理后线粒体活性氧(mtROS)水平;流式细胞术检测细胞凋亡水平。结果 Res抑制RA-FLSs增殖,降低细胞内MnSOD表达;荧光倒置显微镜与Western blot检测证实慢病毒可高效感染RA-FLSs,使MnSOD表达上调和抑制。与对照组相比,在同种因素处理下,MnSOD过表达组mtROS水平降低,凋亡细胞减少。而MnSOD干扰组则呈现相反的结果。结论 Res可通过抑制MnSOD表达来上调mtROS水平,从而促进RA-FLSs凋亡。

Resveratrol mediates apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by down-regulating MnSOD

Aim To investigate the role of manganese superoxide dismutase(MSOD) protein expression in the proliferation and apoptosis of fibroblast-like syno- viocyte (RA-FLS) in rheumatoid arthritis mediated by resveratrol(Res).Methods After RA-FLSs was treated with Res,the expression of MnSOD protein was detected by CCK-8 method,and the expression of MnSOD protein was detected by,Western blot.After lentivirus infection with RA-FLSs,8 mg·L^-1 purine mycin was used to screen the infected cells,and three stable cell lines were established:MnSOD overexpression,MnSOD interference and MnSOD control.The mitochondria reactive oxygen species(mtROS) levels of each cell line treated with 5 μmol·L^-1 hydrogen peroxide(H 2O 2) and 200 mol·L^-1 Res were detected by laser confocal microscopy,and cell apoptosis was detected by flow cytometry.Results Res could inhibit the proliferation and the expression of MnSOD in RA-FLSs.The lentivirus-mediated MnSOD regulation could significantly increase or decrease the expression of MnSOD in RA-FLSs.MnSOD over expression could decrease the level of mtROS and apoptosis of the RA-FLSs treated with 5 μmol·L^-1 H 2O 2 and 200 μmol·L^-1 Res.While the MnSOD RNAi showed the opposite results.Conclusions Res may up-regulate mtROS levels by inhibiting the expression of MnSOD,thus promoting the apoptosis of RA-FLSs.