绿原酸调控miR-124/STAT3轴减轻ox-LDL诱导的血管内皮细胞炎症损伤  

Chlorogenic acid regulates miR-124/STAT3 axis to reduce ox-LDL-induced inflammatory injury of vascular endothelial cells

作　　者：杨洋 徐彧 赵艳芳 YANG Yang;XU Yu;ZHAO Yanfang(Department of Cardiology,Qinhuai Medical District,Eastern Theater General Hospital,Nanjing 210000,China)

机构地区：[1]东部战区总医院秦淮医疗区心血管内科,南京210000

出　　处：《中国免疫学杂志》2023年第4期709-714,共6页Chinese Journal of Immunology

基　　金：南京军区医学科技创新课题项目(15MS062)。

摘　　要：目的:探讨绿原酸(CGA)通过调节miR-124/信号转导和转录活化因子3(STAT3)轴对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)炎症损伤的影响。方法:体外培养HUVECs细胞,分为对照组、ox-LDL组、ox-LDL+CGA 25、50、100μmol/L组、ox-LDL+CGA 100μmol/L+NC inhibitor组、ox-LDL+CGA 100μmol/L+miR-124 inhibitor组,ox-LDL浓度均为150μg/ml。RT-qPCR检测细胞中miR-124、STAT3 mRNA水平;CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡率;ELISA检测细胞培养液中TNF-α、IL-1β、IL-6水平;Western blot检测细胞中STAT3、CyclinD1、Bax、Caspase-3蛋白表达水平;双荧光素酶报告基因实验验证miR-124与STAT3的靶向关系。结果:与对照组相比,ox-LDL组HUVECs凋亡率、Bax、Caspase-3蛋白表达、TNF-α、IL-1β、IL-6水平、STAT3 mRNA及蛋白表达水平显著升高,细胞OD值、CyclinD1蛋白表达、miR-124表达水平显著降低(P<0.05);与ox-LDL组比较,ox-LDL+CGA 25、50、100μmol/L组细胞凋亡率、Bax、Caspase-3蛋白表达、TNF-α、IL-1β、IL-6水平、STAT3 mRNA及蛋白表达水平显著降低,细胞OD值、CyclinD1蛋白表达、miR-124表达水平显著升高(P<0.05);抑制miR-124表达可逆转CGA对ox-LDL诱导的HUVECs细胞炎症损伤的减轻作用;双荧光素酶报告基因实验显示miR-124与STAT3存在靶向关系。结论:CGA可减轻ox-LDL诱导的HUVECs炎症损伤,其作用机制与上调miR-124及抑制STAT3表达有关。Objective:To investigate the effect of chlorogenic acid(CGA)on the inflammatory injury of human umbilical vein endothelial cells(HUVECs)induced by oxidized low-density lipoprotein(ox-LDL)via the regulation of miR-124/signal transducer and activator of transcription 3(STAT3)axis.Methods:HUVECs cells were cultured in vitro and divided into control group,ox-LDL group,ox-LDL+CGA 25,50,100μmol/L groups,ox-LDL+CGA 100μmol/L+NC inhibitor group and ox-LDL+CGA 100μmol/L+miR-124 inhibitor group,the ox-LDL concentration was 150μg/ml.RT-qPCR method was used to detect the levels of miR-124 and STAT3 mRNA in cells;CCK-8 method was used to detect cell proliferation;flow cytometry was used to detect cell apoptosis rate;ELISA was used to detect the levels of TNF-α,IL-1βand IL-6 in cell culture fluid;Western blot method was used to detect the expression levels of STAT3,CyclinD1,Bax and Caspase-3 proteins in cells;dual luciferase reporter gene experiment was used to verify the targeting relationship between miR-124 and STAT3.Results:Compared with the control group,the HUVECs apoptosis rate,Bax and Caspase-3 protein expressions,TNF-α,IL-1βand IL-6 levels,STAT3 mRNA and protein expression levels in the ox-LDL group were significantly increased,the cell OD value,CyclinD1 protein expression and miR-124 expression level were significantly reduced(P<0.05);com‐pared with the ox-LDL group,the HUVECs cell apoptosis rate,Bax,Caspase-3 protein expression,TNF-α,IL-1β,IL-6 levels,STAT3 mRNA and protein expression levels in the ox-LDL+CGA 25,50,100μmol/L groups were significantly reduced,the cell OD value,CyclinD1 protein expression and miR-124 expression level were significantly increased(P<0.05);inhibiting the expression of miR-124 could reverse the effect of CGA on reducing the inflammatory damage of HUVECs induced by ox-LDL;the dual luciferase reporter gene experiment showed that miR-124 and STAT3 had a targeting relationship.Conclusion:CGA can alleviate the inflamma‐tory injury of HUVECs induced by ox-LDL,and its mechanism is

关 键 词：绿原酸 氧化低密度脂蛋白 人血管内皮细胞 炎症损伤 微小RNA-124/信号转导和转录活化因子3轴 

分 类 号：R541.[医药卫生—心血管疾病]