【摘要】 目的 探讨沉默miR-126对关节软骨细胞增殖、凋亡和炎症反应的影响及其机制。方法 体外传代培养关节软骨细胞HC-a。取传3代、对数生长期、生长状态良好的HC-a细胞，随机分为对照组、IL-1β组、IL-1β+miR-NC组、IL-1β+miR-126 inhibitor组，IL-1β+miR-NC组和IL-1β+miR-126 inhibitor组分别转染miR-NC和miR-126 inhibitor；除对照组外，其余三组均予10 ng/mL IL-1β诱导骨关节炎细胞模型。收集各组细胞，采用RT-qPCR法检测miR-126表达，采用CCK-8法检测细胞增殖活性，采用流式细胞术检测细胞凋亡率，采用ELISA法检测培养上清液炎症因子TNF-α、IL-6、IFN-γ，采用Western blotting法检测SIRT1、Caspase-3蛋白表达。结果 与对照组比较，IL-1β组miR-126相对表达量升高，细胞增殖活性下降，细胞凋亡率升高，培养上清液TNF-α、IL-6、IFN-γ水平升高，SIRT1蛋白相对表达量降低，Caspase-3蛋白相对表达量升高（P均<0.05）；与IL-1β+miR-NC组比较，IL-1β+miR-126 inhibitor组miR-126相对表达量降低，细胞增殖活性升高，细胞凋亡率降低，培养上清液TNF-α、IL-6、IFN-γ水平下降，SIRT1蛋白相对表达量升高，Caspase-3蛋白相对表达量降低（P均<0.05）；而IL-1β组和IL-1β+miR-NC组上述指标比较差异均无统计学意义（P均>0.05）。结论 沉默miR-126能够促进关节软骨细胞增殖并抑制其凋亡和炎症反应，其机制可能与调控SIRT1蛋白表达有关。更多还原

【Abstract】 Objective To investigate the effects of silencing miR-126 on proliferation, apoptosis, and inflammatory response of osteoarticular chondrocytes and their mechanism. Methods Human osteoarticular chondrocytes HC-a were cultured in vitro. HC-a cells in the third generation, which were in the logarithmic growth phase and had good growth status, were randomly divided into the control group, IL-1β group, IL-1β + miR-NC group, and IL-1β + miR-126 inhibitor group, respectively. Cells in the IL-1β + miR-NC group and IL-1β + miR-126 inhibitor group were transfected with miR-NC and miR-126 inhibitor, respectively. Except for the control group, the other three groups were treated with 10 ng/mL IL-1β induction to construct the cell models of osteoarthritis(OA). The cells of each group were collected, and the expression of miR-126 was detected by RT-qPCR, cell proliferation activity was detected by CCK-8, apoptosis rate was detected by flow cytometry, and inflammatory factors TNF-α, IL-6 and IFN-γ in the culture supernatant were detected by ELISA.The expression levels of SIRT1 and Caspase-3 proteins were detected by Western blotting. Results Compared with the control group, the relative expression of miR-126 increased, the cell proliferation activity decreased, apoptosis rate increased, the levels of TNF-α, IL-6 and IFN-γ in culture supernatance increased, the relative expression of SIRT1 protein decreased, and the relative expression of Caspase-3 protein increased in the IL-1β group(all P<0. 05). Compared with the IL-1β + miR-NC group, the relative expression decreased, cell proliferation activity increased, apoptosis rate decreased, the levels of TNF-α, IL-6 and IFN-γ in culture supernatant decreased, the relative expression of SIRT1 protein increased, and the relative expression of Caspase-3 protein decreased in the miR-126 inhibitor group(all P<0. 05).There were no significant differences in the above indexes between IL-1β group and IL-1β + miR-NC group(all P>0. 05).Conclusion Silencing miR-126 can promote the proliferation of OA cells and inhibit apoptosis and inflammation, which may be related to the regulation of SIRT1 protein expression.更多还原