高良姜素通过下调lncRNA H19的表达抑制ox-LDL诱导的人源 正常主动脉内皮细胞血管生成活性  

Galangin inhibits oxidized low-density lipoprotein-induced angiogenic activity in human aortic endothelial cells by downregulating lncRNA H19

作　　者：罗瑞 田龙海[1] 杨永曜[1] LUO Rui;TIAN Longhai;YANG Yongyao(Department of Cardiovascular Medicine,Guizhou Provincial People's Hospital,Guiyang 550002,China)

机构地区：[1]贵州省人民医院心血管内科,贵州贵阳550002

出　　处：《南方医科大学学报》2024年第1期52-59,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(82260094);贵州省科学技术基金(黔科合基础-ZK[2021]一般355)。

摘　　要：目的探索高良姜素对ox-LDL诱导的人源正常主动脉内皮细胞(HAECs)的影响及其机制。方法使用不同浓度(0、10、20、40、80μmol/L)的高良姜素孵育HAECs24h后,通过MTT检测细胞活性。以5mg/mLox-LDL诱导的HAECs为模型,使用20、40μmol/L高良姜素孵育24h。为观察高良姜素的作用机制,在HAECs中过表达lncRNAH19,使用40μmol/L高良姜素孵育24 h。通过qRT-PCR检测lncRNAH19的水平,划痕实验和血管形成实验用于观察迁移和管形成能力,使用流式细胞术检测ROS水平。通过Western blot检测VEGFA、MMP-2、MMP-9的蛋白水平。结果低浓度(0、10、20、40μmol/L)的高良姜素对细胞无明显毒性(P>0.05),而80μmol/L高良姜素会抑制HAECs细胞的活性(P<0.01)。ox-LDL诱导的HAECs中lncRNAH19的表达水平增加,高良姜素能降低ox-LDL诱导的HAECs的lncRNAH19的表达水平、迁移能力和血管形成能力,ROS水平以及VEGFA、MMP-2、MMP-9的蛋白水平(P<0.01)。但是,高良姜素的这些生物学作用均能被过表达lncRNAH19逆转(P<0.01)。结论高良姜素可能通过抑制lncRNAH19的表达以降低ox-LDL诱导的HAECs的迁移、氧化应激和血管形成能力来发挥治疗动脉粥样硬化的潜力。Objective To investigate the effects of galangin on angiogenic activity of oxidized low-density lipoprotein(ox-LDL)-induced human aortic endothelial cells(HAECs)and explore the underlying mechanisms.Methods HAECs incubated with 10,20,40,and 80μmol/L galangin for 24 h were assessed for cell viability changes using MTT assay to determine the cytotoxicity of galangin.HAECs treated with 5 mg/mL ox-LDL and incubated with 20 and 40μmol/L galangin for 24 h,and the cells overexpressing lncRNA H19 and incubated with 40μmol/L galangin for 24 h were examined for lncRNA H19 level with qRT-PCR.The migration and tube formation capacity of the cells were observed using scratch assay and angiogenesis assay,and ROS levels in the cells were detected with flow cytometry.The protein expression levels of VEGFA,MMP-2 and MMP-9 in the treated cells were detected with Western blotting.Results Galangin at 10,20,or 40μmol/L produced no obvious toxicity(P>0.05),whereas 80μmol/L galangin significantly inhibited the viability of HAECs(P<0.01).Treatment with ox-LDL significantly increased the expression of lncRNA H19 in HAECs.Galangin significantly lowered lncRNA H19 expression in ox-LDL-induced HAECs,suppressed cell migration,angiogenesis and ROS production level,and reduced the protein levels of VEGFA,MMP-2 and MMP-9(P<0.01).The effects of galangin were blocked by overexpression of lncRNA H19 in the cardiomyocytes.Conclusion The therapeutic effect of galangin for atherosclerosis is mediated by inhibiting lncRNA H19 expression to reduce ox-LDL-induced migration,oxidative stress,and angiogenesis of HAECs.

关 键 词：血管生成 人源正常主动脉内皮细胞 高良姜素 lncRNAH19 

分 类 号：R285[医药卫生—中药学]