下调长链非编码RNA XIST靶向miR-124/NF-κB轴缓解IL-1β诱导的软骨细胞凋亡

Downregulation of Long Non-coding RNA XIST Alleviates IL-1β-induced Chondrocyte Apoptosis by Targeting miR-124/NF-κB Axis

目的探究长链非编码RNA(long non-coding RNA,lncRNA)X染色体失活特异性转录本基因(X inactive specific transcript,XIST)对白细胞介素-1β(interleukin-1β,IL-1β)诱导的软骨细胞凋亡的影响及其作用机制。方法生物信息学预测XIST和miR-124靶向关系并通过双荧光素酶法进行验证;实验设置正常组、IL-1β组、si-NC组、si-XIST组、miR-124-NC组、miR-124 mimics组、si-NC+inhibitor-NC组、si-XIST+inhibitor-NC组、si-NC+miR-124 inhibitor组、si-XIST+miR-124 inhibitor组。qRT-PCR检测细胞中XIST和miR-124表达;蛋白质印迹检测核因子κB P65(nuclear factor kappa B P65,NF-κB P65)蛋白表达;流式细胞术检测细胞凋亡。结果lncRNA XIST和miR-124间存在靶向关系;相较于正常组,IL-1β诱导的软骨细胞中lncRNA XIST表达水平、NF-κB P65蛋白表达和细胞凋亡率显著升高,miR-124表达水平显著降低(P<0.05);相较于IL-1β组,si-XIST组和miR-124 mimics组NF-κB P65蛋白表达和细胞凋亡率显著降低(P<0.05);相较于si-NC+inhibitor-NC组,si-XIST+inhibitor-NC组NF-κB P65蛋白表达和细胞凋亡率显著降低,与si-NC+miR-124 inhibitor组比较,si-XIST+miR-124 inhibitor组NF-κB P65蛋白表达和细胞凋亡率显著降低(P<0.05)。结论下调lncRNA XIST可靶向调节miR-124/NF-κB轴,从而缓解IL-1β诱导的软骨细胞凋亡。

Objective To explore the effect of long non-coding RNA(lncRNA)X inactive specific transcript(XIST)on interleukin-1β(IL-1β)-induced chondrocyte apoptosis and its mechanism.Methods The targeting relationship between XIST and miR-124 was predicted by bioinformatics and verified by dual luciferase reporter gene assay.Ten experimental groups were set as follows:normal group,IL-1βgroup,si-NC group,si-XIST group,miR-124-NC group,miR-124 mimics group,si-NC+inhibitor-NC group,si-XIST+inhibitor-NC group,si-NC+miR-124 inhibitor group,si-XIST+miR-124 inhibitor group.qRT-PCR was used to detect the expression levels of XIST and miR-124 in cells.Western blotting was used to detect the expression level of nuclear factor kappa B P65(NF-κB P65)protein.Flow cytometry was used to detect cell apoptosis.Results There was a targeting relationship between lncRNA XIST and miR-124.The expression levels of lncRNA XIST and NF-κB P65 protein and the apoptosis rate in the IL-1β-induced chondrocytes were increased,while the expression level of miR-124 was decreased when compared with those in the normal group(P<0.05).The expression level of NF-κB P65 protein and the apoptosis rate in the si-XIST group and the miR-124 mimics group were reduced when compared with those in the IL-1βgroup(P<0.05).The expression level of NF-κB P65 protein and the cell apoptosis rate in the si-XIST+inhibitor-NC group were reduced when compared with those in the si-NC+inhibitor-NC group.The expression level of NF-κB P65 protein and the apoptosis rate in the si-XIST+miR-124 inhibitor group were decreased when compared with those in the si-NC+miR-124 inhibitor group,(P<0.05).Conclusion Down-regulation of lncRNA XIST can target and regulate the miR-124/NF-κB axis,thereby alleviating the apoptosis of chondrocytes induced by IL-1β.