丹参酮ⅡA磺酸钠对LPS引起的HUVEC功能异常和凋亡调控作用的研究
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金(81770450)


Research on the regulatory effects of sodium tanshinone ⅡA sulphonate on dysfunction and apoptosis of HUVEC induced by LPS
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:研究丹参酮ⅡA磺酸钠(sodium tanshinone ⅡA sulphonate,STS)对脂多糖(lipopolysaccharide,LPS)引起的人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)功能异常和凋亡的调控作用。方法:将第6~8代处于对数生长期的HUVEC按如下分组给予处理:DMEM组、LPS(1.0 μg/mL)处理组、高浓度(50.0 μg/mL)STS预处理组、中浓度(25.0 μg/mL)STS预处理组和低浓度(12.5 μg/mL)STS预处理组。其中,各STS预处理组均先以对应浓度的STS预处理HUVEC 2 h,再加入1 μg/mL LPS进行刺激。处理24 h后,CCK-8 法测定细胞活力;流式细胞术检测细胞增殖;ELISA 法检测细胞培养上清中炎症因子白介素1β(interleukin-1β,IL-1β)的蛋白浓度;Western blot法检测细胞裂解液中IL-1β和凋亡相关分子cleaved caspase-9和cleaved caspase-3的表达,以及细胞核蛋白中核因子κB-p65(nuclear factor-κB-p65,NF-κB-p65)的水平。此外,划痕实验检测HUVEC的迁移能力;光镜观察细胞形态变化;DAPI染色显示细胞核染色质的改变;Annexin V/PI双染法检测HUVEC的凋亡情况。结果:与DMEM相比,LPS处理导致HUVEC活力和迁移能力减低,但促进其增殖;IL-1β和NF-κB p65蛋白水平升高;cleaved caspase-3和cleaved caspase-9表达增高并伴有凋亡水平提高(P<0.05)。STS预处理能够以浓度依赖的方式部分或完全逆转LPS引起的上述现象(P<0.05)。结论:STS能够通过浓度依赖的方式抑制LPS引起的HUVEC功能异常和凋亡,对血管内皮起到保护作用。

    Abstract:

    Objective:This study aims to investigate the effects of sodium tanshinone type ⅡA sulphonate(STS)on lipopolysaccharide(LPS)-induced dysfunction and apoptosis of human umbilical vein endothelial cells(HUVEC). Methods:Cultured HUVEC of 6-8 generation were grouped and stimulated as follows:DMEM group,LPS(1.0 μg/mL) group,high concentration(50.0 μg/mL)STS pre-treated group,medium concentration(25.0 μg/mL)STS pre-treated group,low concentration(12.5 μg/mL) STS pre-treated group. For STS pre-treated groups,HUVEC were pre-treated with corresponding concentrations of STS for 2 h,followed by the stimulation with 1 μg/mL LPS. After 24 h stimulation,cell viability was determined by CCK-8 method. Cell proliferation was detected by flow cytometry. Protein level of inflammatory cytokine interleukin-1 beta(IL-1β) in culture supernatant was determined by ELISA. Western blotting was employed for the detection of protein levels of intracellular IL-1β,cleaved caspase-3 and cleaved caspase-9,and nucleus-translocated nuclear factor-κB-p65(NF-κB-p65). Then,migration ability of HUVEC was investigated by wound-healing test. The changes of cell and chromatin morphology in HUVEC were observed by microscopy,and the apoptosis of HUVEC was detected by Annexin V/PI staining. Results:Compared with DMEM,LPS treatment reduced viability and migration capacity of HUVEC,whereas promoted their proliferation. The protein levels of IL-1β,NF-κB-p65,cleaved caspase-3 and cleaved caspase-9 in HUVEC,as well as their apoptosis level were increased after LPS stimulation(P < 0.05). However,STS can partially reverse the described effects of LPS in a dose-dependent manner(P < 0.05). Conclusion:STS inhibits LPS-induced dysfunction and apoptosis in HUVEC,thus exerts protective effects on vascular endothelium.

    参考文献
    相似文献
    引证文献
引用本文

尹超云,潘雅妮,刘 彬,车 媛,王中群,陶 政.丹参酮ⅡA磺酸钠对LPS引起的HUVEC功能异常和凋亡调控作用的研究[J].南京医科大学学报(自然科学版),2020,(11):1590-1596

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2019-09-07
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2020-12-04
  • 出版日期:
通知关闭
郑重声明