摘要
[目的]观察大气细颗粒物PM_(2.5)对体外培养血管内皮细胞坏死性凋亡的影响。[方法]以人微血管内皮细胞株(HMEC-1)为研究对象,应用系列浓度PM_(2.5)处理24 h后,CCK-8检测HMEC-1细胞活力,试剂盒法检测培养上清液乳酸脱氢酶(LDH)的活性,Western blot法检测坏死性凋亡相关蛋白RIPK3和MLKL的表达及磷酸化水平,Transwell法检测HMEC-1细胞通透性改变。采用坏死性凋亡抑制剂(Ne-crostatin-1)处理后,观察PM_(2.5)对HMEC-1坏死性凋亡、细胞损伤和通透性的影响。[结果]PM_(2.5)可呈剂量依赖性抑制HMEC-1的增殖,并增加培养上清液中LDH活性、RIPK3和MLKL的蛋白表达以及其磷酸化水平。Transwell检测结果表明PM_(2.5)可增加单层血管内皮细胞通透性,而坏死性凋亡抑制剂Necrostatin-1预处理可显著性抑制PM_(2.5)应激下血管内皮细胞RIPK3和MLKL的蛋白表达及磷酸化水平,降低上清液LDH活性和单层血管内皮细胞通透性。[结论]PM_(2.5)可诱导HMEC-1坏死性凋亡,损伤血管内皮完整性。
[Objective]To investigate the effect of atmospheric fine particles(particulate matter)PMs on necroptosis of human vascular endothelial cells.[Methods]After 24 hours of treatment with a series of concen-trations of PM_(2.5),CCK-8 was used to detect cell viability in HMEC-1 cells(human microvascular endothelial cell line);and the culture supernatant was used to deteet the level of lactate dehydrogenase(LDH)produced by HMEC-1 cells.Western blot method was applied to detect the expression and phosphorylation level of nee-roptosis-related proteins RIPK3 and MLKL.The permeability changes of vascular endothelial cells were meas-ured by trans well method.After treatment with necrostatin-l,an inhibitor of necroptosis,the efects of PM_(2.5)on necroptosis.cell damage and permeability of vascular endothelial cells were observed and evaulated.[Re-suls]PM_(2.5)inhibited the proliferation of human microvascular endothelial cells in a dose dependent manner.It increased the LDH activity in the culture supernatant and upregulated protein expression and phosphorylation levels of RIPK3 and MLKL.The result of trans-well assay showed that PM_(2.5)increased the permeability of monolayer vascular endothelial cells.The pretreatment of necrostatin-1,an inhibitor of necroptosis,signifi-cantly inhibited the protein expression and phosphorylation levels of RIPK3 and MLKL in vascular endothelial cells under PM_(2.5)stimulation,and reduced the activity of LDH in the supernatant and the permeability of mon-olayer vascular endothelial cells.[Conclusion]PM_(2.5)induces necroptosis of vascular endothelial cells and im-pairs the integrity of vascular endothelium.
作者
黄一丹
马丽
周祎琦
董良
黄春波
HUANG Yi-dan;MA Li;ZHOU Yi-qi(Department of Anesthesiology,Liuzhou Municipal Peoples Hospital,Liuzhou Guangxi 545006)
出处
《医学临床研究》
CAS
2021年第4期488-492,共5页
Journal of Clinical Research
基金
国家自然科学基金资助项目(编号:81760019)。
