上调miR-31表达对AGEs诱导的软骨细胞损伤的影响  

Effect of up regulation of miR-31 expression on AGEs induced chondrocyte injury

作　　者：刘亚东 李刚 卢小伟 LIU Ya-Dong;LI Gang;LU Xiao-Wei(Sinopharm Dongfeng General Hospital of Shiyan City,Hubei Province,Shiyan 442000,China)

机构地区：[1]湖北省十堰市国药东风总医院,十堰442000

出　　处：《中国免疫学杂志》2021年第9期1058-1062,共5页Chinese Journal of Immunology

摘　　要：目的:探讨上调miR-31表达对晚期糖基化终末产物(AGEs)诱导的软骨细胞凋亡、炎症反应和氧化应激的影响。方法:将体外培养的大鼠软骨细胞分为对照组、AGEs组、AGEs+miR-NC组和AGEs+miR-31组,采用RT-PCR法检测软骨细胞中miR-31表达,MTT法检测软骨细胞活力,流式细胞仪检测细胞凋亡率,比色法检测细胞Caspase-3活性,Western blot检测细胞中Bcl-2和Bax蛋白表达水平,试剂盒检测细胞上清液中炎症因子IL-1β、TNF-α和氧化应激因子SOD、MDA含量。结果:与对照组比较,AGEs组细胞存活率、细胞上清液中SOD含量和细胞中Bcl-2蛋白表达水平显著降低,而细胞上清液中IL-1β、TNF-α、MDA含量和细胞凋亡率、Caspase-3活性以及Bax蛋白表达水平均显著升高(P<0.05);与AGEs组比较,AGEs+miR-NC组中各指标无显著差异(P>0.05);与AGEs组或AGEs+miR-NC组比较,AGEs+miR-31组细胞存活率、细胞上清液中SOD含量和细胞中Bcl-2蛋白表达水平均显著升高,而细胞上清液中IL-1β、TNF-α、MDA含量、细胞凋亡率、Caspase-3活性以及Bax蛋白表达水平均显著降低(P<0.05)。结论:上调miR-31表达可通过抑制AGEs诱导的细胞凋亡、炎症反应和氧化应激起到保护软骨细胞损伤的作用。Objective:To investigate the effect of up regulation of miR-31 expression on chondrocyte apoptosis,inflammatory response and oxidative stress induced by advanced glycation end products(AGEs).Methods:The chondrocytes cultured in vitro were divided into four groups:control group,AGEs group,AGEs+miR-NC group and AGEs+miR-31 group.The expression of miR-31 in chondrocytes was detected by RT-PCR,the activity of chondrocytes was detected by MTT,the apoptosis rate was detected by flow cytometry,the caspase-3 activity was detected by colorimetry,the expressions of Bcl-2 and Bax proteins were detected by Western blot,and the contents of IL-1β,TNF-α,SOD and MDA in the supernatantthe cells were detected by kit.Results:compared with the control group,the cell survival rate,SOD content in the supernatant and Bcl-2 protein expression level were significantly reduced,while the IL-1β,TNF-α,MDA content in the supernatant,apoptosis rate,Caspase-3 activity and Bax protein expression level in AGEs group were significantly increased(P<0.05);compared with AGEs group,the indexes of AGEs+miR-NC group were not significantly different(P>0.05);compared with AGEs group or AGEs+miR-NC group,the cell survival rate,SOD content in supernatant and Bcl-2 protein expression level in AGEs+miR-31 group were significantly increased,while the IL-1β,TNF-α,MDA content in supernatant,apoptosis rate,Caspase-3 activity and Bax protein expression level were significantly decreased(P<0.05).Conclusion:Up regulation of miR-31 can protect chondrocytes from injury by inhibiting AGEs induced apoptosis,inflammation and oxidation.

关 键 词：软骨细胞 miR-31 炎症 氧化应激 细胞凋亡 

分 类 号：R684.3[医药卫生—骨科学]