摘要
目的探讨长链非编码RNA(lncRNA)NORAD对氧化型低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞功能障碍的影响及可能的分子机制。方法将人脐静脉内皮细胞分为阴性对照组、实验组1、实验组2和实验组3,分别用不同浓度(0、50、100、200μg/mL)ox-LDL处理。将人脐静脉内皮细胞分为阴性对照组、ox-LDL组、ox-LDL+shNC组和ox-LDL+shNORAD组,后两组分别转染shNC和shNORAD,且后三组用200μg/mL的ox-LDL处理。采用实时荧光定量PCR检测lncRNA NORAD表达,采用Western blotting法检测NF-κB通路的关键因子NF-κB p65、p-IκBα表达,采用ELISA法检测白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)及细胞间黏附分子1(ICAM-1)。结果随着ox-LDL浓度的增加,lncRNA NORAD与NF-κB p65、p-IκBα的表达均升高(P均<0.05)。选择200μg/mL的ox-LDL进行下一步实验。与ox-LDL+shNC组相比,ox-LDL+shNORAD组lncRNA NORAD、NF-κB p65、p-IκBα表达及IL-6、TNF-α、ICAM-1水平下降(P均<0.05)。结论下调lncRNA NORAD表达可降低ox-LDL诱导的内皮细胞分泌趋化因子与炎症因子的能力,其作用机制可能与对NF-κB通路的抑制有关。
Objective To investigate the effect of long non-coding RNA NORAD(lncRNA NORAD)on the dysfunction of human umbilical vein endothelial cells(HUVECs)induced by oxidized low-density lipoprotein(ox-LDL)and its possible molecular mechanism.Methods HUVECs were divided into the negative control group,experimental group 1,experimental group 2,and experimental group 3.The corresponding cells were treated with different concentrations of ox-LDL(0,50,100 and 200μg/mL).HUVECs were divided into the negative control group,ox-LDL group,ox-LDL+shNC group,and ox-LDL+shNORAD group.Cells in the ox-LDL+shNC group and the ox-LDL+shNORAD group were transfected with shNC and shNORAD,respectively.Then the cells in the latter three groups were treated with 200μg/mL ox-LDL.The expression of lncRNA NORAD was detected by real-time quantitative PCR(qPCR).The expression levels of key factors NF-κB p65 and p-IκBαin NF-κB signaling pathway were detected by Western blotting.The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and intercellular adhesion molecule-1(ICAM-1)were detected by ELISA.Results With the increase of ox-LDL concentrations,the expression of lncRNA NORAD,NF-κB p65 and p-IκBαincreased(all P<0.05).The ox-LDL(200μg/mL)was selected for the next experiment.Compared with the ox-LDL+shNC group,the expression of lncRNA NORAD,NF-κB p65 and p-IκBα,and the levels of 1L-6,TNF-α,and ICAM-1 in the ox-LDL+shNORAD group decreased(all P<0.05).Conclusion Down-regulation of lncRNA NORAD expression can reduce the ability of endothelial cells to secrete chemokines and inflammatory cytokines under oxidative stress,and its mechanism may be related to the inhibition of NF-κB pathway.
作者
赵晓雪
张冰玉
刘宇翔
ZHAO Xiaoxue;ZHANG Bingyu;LIU Yuxiang(Comprehensive Geriatrics Ward 3,Liaoning Jinqiu Hospital,Shenyang 110016,China)
出处
《山东医药》
CAS
2021年第19期48-52,共5页
Shandong Medical Journal
