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
罗瑞 1田龙海 1杨永曜1
作者信息
- 1. 贵州省人民医院心血管内科,贵州 贵阳 550002
- 折叠
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、80 μmol/L)的高良姜素孵育HAECs 24 h后,通过MTT检测细胞活性.以5 mg/mL ox-LDL诱导的HAECs为模型,使用20、40 μmol/L高良姜素孵育24 h.为观察高良姜素的作用机制,在HAECs中过表达lncRNA H19,使用40 μmol/L高良姜素孵育24 h.通过qRT-PCR检测lncRNA H19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、40 μmol/L)的高良姜素对细胞无明显毒性(P>0.05),而80 μmol/L高良姜素会抑制HAECs细胞的活性(P<0.01).ox-LDL诱导的HAECs中lncRNA H19的表达水平增加,高良姜素能降低ox-LDL诱导的HAECs的lncRNA H19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lncRNA H19逆转(P<0.01).结论 高良姜素可能通过抑制lncRNA H19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.
Abstract
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 80 μmol/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 40 μmol/L galangin for 24 h,and the cells overexpressing lncRNA H19 and incubated with 40 μmol/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 40 μmol/L produced no obvious toxicity(P>0.05),whereas 80 μmol/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/lncRNA/H19Key words
angiogenesis/human aortic endothelial cells/galangin/lncRNA H19引用本文复制引用
基金项目
国家自然科学基金(82260094)
贵州省科技计划(82260094)
出版年
2024
国家科技期刊平台
NSTL
万方数据