TNF-α对人脐静脉内皮细胞活性及PD-L1蛋白表达的影响  

The effects of recombinant human TNF-α on the expression of programmed death-1 ligand-1 and proliferation of human umbilical vein endothelial cells

作　　者：吕自明[1] 刘映峰[1] 缪绯[1] 龙日[1] 刘芃[1] 刘磊[1] 

机构地区：[1]南方医科大学附属珠江医院心血管内科,广东广州510280

出　　处：《热带医学杂志》2013年第6期691-694,F0002,共5页Journal of Tropical Medicine

基　　金：广东省自然科学基金(10151051501000038)

摘　　要：目的研究外源性肿瘤坏死因子α(TNF-α)对原代人脐静脉内皮细胞(HUVECs)活性及程序性死亡因子配体PD-L1蛋白表达的影响,探讨动脉粥样硬化的免疫机制。方法体外培养HUVECs,加入不同浓度TNF-α持续刺激48h,CCK-8法检测450nm波长处吸光度值(OD值),计算细胞生长存活率。选同代细胞重复培养并干预,Westernblot方法检测细胞PD-L1蛋白表达。使用SB203580阻断p38MAPK通路后再用TNF-α干预HUVECs,比较各组细胞PD-L1蛋白表达。结果实验组HUVECs在450nm波长处的吸光度值较空白对照组明显降低(P<0.05),细胞存活率依次下降。实验组细胞PD-L1蛋白表达随TNF-α浓度增高逐渐降低(P<0.05)。TNF-α单独刺激组PD-L1蛋白表达较TNF-α与SB203580共刺激组明显降低(P<0.05),SB203580阻断效应明显。结论 TNF-α可明显抑制HUVECs细胞活性,并降低PD-L1蛋白表达,p38MAPK通路可能参与了这一过程。Objective This study aims to （i） investigate the effect of TNF-α on the expression of PD-L1 and on the proliferation of human umbilical vein endothelial cells （HUVECs）, and （ii） study the immunopathogenesis of atherosclerosis. Methods HUVECs were incubated with various concentrations of TNF-α for 48 hours. Cell viability and proliferation were determined by CCK-8 assay. The protein of PD-L1 was analyzed by Western blotting. p38MAPK inhibitor SB203580 was added before TNF-α treatment. Results As the concentration of TNF-α increased from 5 ng / ml to 80 ng / ml, the OD value at 450 nm in the CCK-8 assay decreased significantly （P〈0.05）. The expression of PD-L1 protein was suppressed by TNF-α significantly when compared with the control group （P〈0.05）. The level of PD-L1 protein in TNF-α-treated group was lower than the normal group and TNF-α ＋ SB203580 group （P〈0.05）. Conclusion TNF-α was found to suppress the expression of PD-L1 and inhibit the proliferation of HUVECs. p38MAPK pathway may play a role in this process.

关 键 词：肿瘤坏死因子Α PD-L1 P38MAPK 脐静脉内皮细胞 

分 类 号：R541.4[医药卫生—心血管疾病]