miR-23a对软骨细胞增殖、分化和氧化应激的影响机制

Mechanism of miR-23a on chondrocyte proliferation,differentiation and oxidative stress

[目的]探究miR-23a调控AMPK/SIRT1通路对人退变关节软骨细胞增殖、分化和氧化应激的影响。[方法]人退变关节软骨分为对照组、miR-23a mimic组、miR-23a inhibitor组。通过对以上三组细胞转染miR-23a NC、miR-23a mimic和miR-23a inhibitor转染来干扰细胞中miR-23a的表达水平。转染完成后,分别通过CCK-8、ELISA和Western Blot实验检测各组细胞增殖、氧化应激水平、分化能力以及AMPK/SIRT1信号通路的表达水平。[结果]三组细胞的各项比较差异显著(P<0.05)。与对照组相比,miR-23a mimic组的miR-23a、骨形成蛋白(BMP)2、BMP4、MDA水平显著升高(P<0.05),而OD值、SOD、AMPK/SIRT1通路水平显著降低(P<0.05)。与对照组相比,miR-23a inhibitor组的miR-23a、BMP2、BMP4、MDA水平显著降低(P<0.05),而OD值、SOD、AMPK/SIRT1通路水平显著升高(P<0.05)。[结论]OA患者的软骨组织中的miR-23a表达水平显著升高,miR-23a在退化的软骨中可抑制软骨细胞增殖,参与软骨分化,并抑制AMPK/SIRT1通路。

[Objective]To explore the effect of miR-23a regulating AMPK/SIRT1 pathway on the proliferation,differentiation and oxidative stress of human degenerated articular chondrocytes.[Method]Human degenerated articular cartilage was divided into control group,miR-23a mimic group and miR-23a inhibitor group.The above three groups of cells were transfected with miR-23a NC,miR-23a mimic and miR-23a inhibitor to interfere the expression level of miR-23a in cells.After transfection,CCK-8,ELISA and Western Blot experiments were used to detect the cell proliferation,oxidative stress level,differentiation ability and the expression level of AMPK/SIRT1 signaling pathway in each group.[Result]There were significant differences among the three groups of cells(P<0.05).Compared with the control group,the levels of miR-23a,bone morphogenetic protein(BMP)2,BMP4 and MDA in the miR-23a mimic group were significantly increased(P<0.05),while the OD value,SOD,and AMPK/SIRT1 pathway levels were significantly decreased(P<0.05).Compared with the control group,the levels of miR-23a,BMP2,BMP4 and MDA in the miR-23a inhibitor group were significantly decreased(P<0.05),while the OD value,SOD,and AMPK/SIRT1 pathway levels were significantly higher than those in the control group(P<0.05).[Conclusion]The expression level of miR-23a in cartilage tissue of OA patients was significantly increased,and miR-23a could inhibit chondrocyte proliferation,participate in cartilage differentiation,and inhibit the AMPK/SIRT1 pathway in degenerated cartilage.