miR-21对类风湿关节炎滑膜成纤维细胞增殖与凋亡的影响

Effects of miR-21 on the proliferation and apoptosis of synovial fibroblasts in rheumatoid arthritis

目的 探讨miR-21对类风湿关节炎(RA)滑膜成纤维细胞(FLS)增殖与凋亡的影响。方法 Realtime PCR方法检测正常和RA-FLS中miR-21表达差异。RA-FLS分成Control组、miR-NC组(转染mimics control)、miR-21组(转染miR-21 mimics)、miR-21+IGF-1组(转染miR-21 mimics, PI3K/Akt信号通路特异性激活剂IGF-1处理),CCK-8实验分析细胞增殖活性变化,流式细胞术分析细胞凋亡水平变化,Western blot方法分析细胞中C-Caspase-3、Bax、Bcl-2、PI3K、p-PI3K、Akt、p-Akt蛋白表达变化。结果 与正常-FLS比较,RA-FLS中miR-21表达水平降低(P<0.05)。与Control组、miR-NC组比较,miR-21组RA-FLS细胞中miR-21表达水平升高,细胞增殖活性降低,细胞凋亡率增加,细胞中C-Caspase-3、Bax蛋白表达增多,Bcl-2蛋白表达减少,p-PI3K/PI3K、p-Akt/Akt蛋白水平降低(P<0.05)。与miR-21组比较,miR-21+IGF-1组RA-FLS增殖活性升高,细胞凋亡率降低,细胞中p-PI3K/PI3K、p-Akt/Akt蛋白表达增多,C-Caspase-3、Bax蛋白表达减少,Bcl-2蛋白表达增多(P<0.05)。结论 上调miR-21可抑制RA-FLS增殖,诱导细胞凋亡,机制可能与降低PI3K/Akt信号激活水平有关。

Objective To investigate the effect of miR-21 on the proliferation and apoptosis of rheumatoid arthritis(RA) fibrolast-like synoviocytes(FLS). Methods Realtime PCR method was used to detect the difference of miR-21 expression between normal and RA-FLS. RA-FLS were divided into Control, miR-NC(transfection mimics control), miR-21(transfection miR-21 mimics), miR-21+IGF-1 group(transfection miR-21 mimics, PI3 K/Akt signaling pathway specific treatment with PI3 K/Akt activator IGF-1), CCK-8 experiment to analyze changes in cell proliferation activity, flow cytometry to analyze changes in cell apoptosis, Western blot analysis of C-Caspase-3, Bax, Bcl-2, PI3 K, p-PI3 K, Akt and p-Akt proteins. Results Compared with normal-FLS, the expression level of miR-21 in RA-FLS decreased(P<0.05). Compared with the Control and miR-NC groups, the miR-21 expression level in the RA-FLS cells of the miR-21 group increased, the cell proliferation activity decreased, the apoptosis rate increased, and the expression of C-Caspase-3 and Bax proteins in the cells increased, the expression of Bcl-2 protein decreased, and the protein levels of p-PI3 K/PI3 K and p-Akt/Akt decreased(P<0.05). Compared with the miR-21 group, the proliferation activity of RA-FLS in the miR-21+IGF-1 group was increased, the apoptosis rate was decreased, the expression of p-PI3 K/PI3 K, p-Akt/Akt protein in the cells increased, and C-Caspase-3,Bax protein decreased, and the expression of Bcl-2 protein increased(P<0.05). Conclusion Up-regulation of miR-21 can inhibit the proliferation of RA-FLS and induce cell apoptosis, the mechanism is related to the reduction of PI3 K/Akt signal activation level.