文章摘要
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,2020,(11):1590~1596
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
Research on the regulatory effects of sodium tanshinone ⅡA sulphonate on dysfunction and apoptosis of HUVEC induced by LPS
投稿时间:2019-09-07  
DOI:10.7655/NYDXBNS20201104
中文关键词: 丹参酮ⅡA磺酸钠  人脐静脉内皮细胞  LPS  IL⁃1β  cleaved caspase⁃3  cleaved caspase⁃9
英文关键词: sodium tanshinone ⅡA sulfonate  human umbilical vein endothelial cell  LPS  IL⁃1β  cleased caspase⁃3  cleaved caspase⁃9
基金项目:国家自然科学基金(81770450)
作者单位
尹超云 江苏大学附属医院血管外科江苏 镇江 212002 
潘雅妮 江苏大学附属医院心电图室江苏 镇江 212002 
刘 彬 江苏大学附属医院血管外科江苏 镇江 212002 
车 媛 江苏大学附属医院血管外科江苏 镇江 212002 
王中群 江苏大学附属医院心内科江苏 镇江 212002 
陶 政 江苏大学附属医院血管外科江苏 镇江 212002 
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中文摘要:
      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。
英文摘要:
      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.
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