LncRNA DANCR靶向miR-423-5p调控ox-LDL诱导的血管内皮细胞凋亡、炎症反应

LncRNA DANCR regulates ox-LDL-induced vascular endothelial cell apoptosis and inflammatory response through targeting miR-423-5p

目的:探讨长链非编码RNA(LncRNA)分化拮抗非蛋白编码RNA(DANCR)对氧化低密度脂蛋白(ox-LDL)处理的血管内皮细胞(HUVEC)凋亡、炎症因子分泌的影响和可能机制。方法:50μg/ml的ox-LDL诱导HUVEC。RT-qPCR检测LncRNA DANCR和miR-423-5p表达水平;细胞计数试剂盒(CCK-8)、流式细胞术分别检测细胞活力和凋亡;ELISA试剂盒检测IL-1β、IL-6、TNF-α水平。将LncRNA DANCR过表达载体、miR-423-5p抑制物分别转染HUVEC,检测过表达LncRNA DANCR或抑制miR-423-5p对ox-LDL诱导的HUVEC增殖、凋亡、炎症因子分泌的影响。双荧光素酶报告基因实验和RT-qPCR确定Ln⁃cRNA DANCR与miR-423-5p之间靶向调控作用。结果:ox-LDL诱导后HUVEC中LncRNA DANCR表达、存活率显著降低,miR-423-5p表达、凋亡率、IL-1β、IL-6、TNF-α水平显著升高(P<0.05)。过表达LncRNA DANCR或抑制miR-423-5p表达后,ox-LDL诱导的HUVEC存活率显著升高,凋亡率显著降低,IL-1β、IL-6、TNF-α水平显著降低(P<0.05)。miR-423-5p是LncRNA DANCR的靶基因,LncRNA DANCR负性调控miR-423-5p表达。过表达miR-423-5p能够逆转LncRNA DANCR过表达对ox-LDL诱导的HUVEC存活、凋亡和炎症因子分泌的影响(P<0.05)。结论:LncRNA DANCR能够提高HUVEC存活率,改善ox-LDL诱导的细胞凋亡和炎症反应,其机制与靶向调控miR-423-5p表达有关。

Objective:To explore the effect of long-face non-coding RNA(LncRNA)differentiation antagonizing non-protein coding RNA(DANCR)on the apoptosis and secretion of inflammatory factors in oxidized low-density lipoprotein(ox-LDL)treated vascular endothelial cells(HUVEC)and its possible mechanism.Methods:HUVEC was treated with 50μg/ml ox-LDL.RT-qPCR detected the expression levels of LncRNA DANCR and miR-423-5p;cell counting kit(CCK-8)and flow cytometry detected cell viability and apoptosis;ELISA kit detected the levels of IL-1β,IL-6 and TNF-α.The LncRNA DANCR overexpression vector and miR-423-5p inhibitor were transfected into HUVEC,respectively.And the effects of overexpression of LncRNA DANCR or the inhibition of miR-423-5p on the proliferation,apoptosis,and inflammatory factor secretion of ox-LDL-induced HUVEC were detected by the above methods.Dual luciferase reporter gene experiment and RT-qPCR confirmed the targeted regulation between LncRNA DANCR and miR-423-5p.Results:After ox-LDL induction,the expression of LncRNA DANCR and survival rate in HUVEC were significantly reduced,and the expression of miR-423-5p,apoptosis rate,IL-1β,IL-6 and TNF-αlevels were significantly increased(P<0.05).After overexpressing LncRNA DANCR or inhibiting miR-423-5p expression,the survival rate of HUVEC induced by ox-LDL was significantly increased,the apoptosis rate,and the levels of IL-1β,IL-6 and TNF-αwere significantly reduced(P<0.05).miR-423-5p is the target gene of LncRNA DANCR.LncRNA DANCR negatively regulates the expression of miR-423-5p.Overexpression of miR-423-5p could reverse the effect of LncRNA DANCR overexpression on the survival,apoptosis and inflammation factor secretion of ox-LDL-induced HUVEC(P<0.05).Conclusion:LncRNA DANCR colud increase the survival rate of HUVEC,ameliorate ox-LDLinduced inflammation and apoptosis,and its mechanism is related to the targeted regulation of miR-423-5p expression.