lncRNA SNHG14靶向miR-149-5p调控MIA诱导的骨关节炎模型细胞损伤

LncRNA SNHG14 targeting miR1495p to regulate MIA-induced cell injury in osteoarthritis models

目的探讨长链非编码RNA SNHG14(lncRNA SNHG14)对碘乙酸钠(MIA)诱导的大鼠软骨细胞损伤的影响及分子机制。方法4μmol/L MIA诱导软骨细胞损伤模型,将其分为对照组、MIA组、MIA+si-NC组、MIA+si-SNHG14组、MIA+miR-NC组、MIA+miR-149-5p组、MIA+si-SNHG14+anti-miR-NC组、MIA+si-SNHG14+anti-miR-149-5p组。实时荧光定量PCR(RT-qPCR)检测lncRNA SNHG14和miR-149-5p的表达水平;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测蛋白表达;比色法测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)水平;双荧光素酶报告实验检测lncRNA SNHG14和miR-149-5p的靶向关系。结果与对照组相比,经MIA诱导的软骨细胞损伤中,lncRNA SNHG14表达水平升高,miR-149-5p表达水平降低(P<0.05),细胞凋亡率升高,Bcl-2表达水平与SOD和CAT活性降低,Bax和MDA表达水平升高(P<0.05)。抑制lncRNA SNHG14和过表达miR-149-5p均降低细胞凋亡率,提高Bcl-2表达水平与SOD和CAT活性,降低Bax和MDA表达水平(P<0.05)。lncRNA SNHG14靶向调控miR-149-5p表达,且下调miR-149-5p能逆转抑制lncRNA SNHG14表达对细胞凋亡及氧化应激反应的影响(P<0.05)。结论抑制lncRNA SNHG14通过靶向上调miR-149-5p减轻MIA诱导的软骨细胞损伤。

Objective To investigate the effects of long non-coding RNA SNHG14(lncRNA SNHG14)on rat chondrocyte injury induced by sodium iodoacetate(MIA)and its molecular mechanism.Methods The chondrocyte injury models were induced by 4μmol/L MIA,then,which were divided into control group,MIA group,MIA+si-NC group,MIA+si-SNHG14 group,MIA+miR-NC group,MIA+miR-149-5p group,MIA+si-SNHG14+anti-miR-NC group,and MIA+si-SNHG14+anti-miR-149-5p group.Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expression levels of lncRNA SNHG14 and miR-149-5p;flow cytometry was used to detect cell apoptosis;Western blot was used to detect protein expression;colorimetric method was used to detect superoxide dismutation enzyme(SOD),catalase(CAT)activity and malondialdehyde(MDA)levels;dual luciferase reporter experiment was conducted to detect the targeting relationship between lncRNA SNHG14 and miR-149-5p.Results Compared with the control group,in the chondrocyte injury induced by MIA,the expression levels of lncRNA SNHG14 were increased,while the expression levels of miR 1495p were significantly decreased(P<0.05).And the apoptosis rate was increased,the expression levels of Bcl-2 and SOD and CAT activity were significantly decreased,and the expression levels of Bax and MDA were significantly increased(P<0.05).The inhibition of lncRNA SNHG14 and overexpression of miR-149-5p reduced the apoptosis rate,increased the expression levels of Bcl-2 and the activity of SOD and CAT,and significantly decreased the expression levels of Bax and MDA(P<0.05).lncRNA SNHG14 targeted and regulated the expression of miR-149-5p,and down-regulation of miR-149-5p could reverse the effects of inhibiting the expression of lncRNA SNHG14 on cell apoptosis and oxidative stress(P<0.05).Conclusion The inhibition of lncRNA SNHG14 can relieve MIA-induced chondrocyte damage by targeting up-regulation of miR-149-5p.