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目的:探讨胺碘酮对人脐静脉内皮细胞（HUVEC）的损伤作用及可能的机制。方法:取经过3代培养的HUVEC接种于96孔板，加入0、10、20、30或60 μmol/L胺碘酮培养24 h，采用细胞计数试剂盒8（CCK-8）法检测细胞活力，以0 μmol/L组细胞活力为100%，计算各加药组的相对细胞活力。选择可将细胞活力降至70%左右的胺碘酮浓度用于后续各项实验。采用CCK-8法检测该浓度胺碘酮作用不同时间（6、12、24、36、48 h）对HUVEC活力的影响。以加入该浓度胺碘酮培养的HUVEC为实验组，不加入者为对照组，采用钙依赖性磷脂结合蛋白V-异硫氰酸荧光素/碘化丙啶双染法流式细胞术检测细胞凋亡率；蛋白质印迹法和实时荧光定量聚合酶链反应法分别检测B细胞淋巴瘤2（Bcl-2）、Bcl-2相关X蛋白（Bax）、半胱氨酸蛋白酶3（caspase-3）、白细胞介素10（IL-10）、IL-1β、IL-6、肿瘤坏死因子α（TNF-α）蛋白和mRNA表达水平；2\',7\'-二氯二氢荧光素二乙酸酯荧光探针法检测活性氧（ROS）含量，水溶性四氮唑-1法检测超氧化物歧化酶（SOD）活性，微板法检测还原型谷胱甘肽（GSH）含量。结果:经10、20、30、60 μmol/L胺碘酮作用24 h的各加药组HUVEC活力与对照组（100%）相比分别为（88.82±2.64）%、（74.96±1.75）%、（64.95±2.10）%和（18.57±0.65）%，各加药组与对照组比较以及加药组之间两两比较均n P<0.01。选择30 μmol/L胺碘酮用于后续实验。经30 μmol/L胺碘酮作用6、12、24、36、48 h的各实验组HUVEC活力与对照组（100%）相比分别为（90.19±1.88）%、（82.81±2.51）%、（75.33±1.37）%、（65.76±1.85）%和（47.01±3.29）%，各实验组与对照组比较以及实验组之间两两比较均n P<0.01。实验组细胞凋亡率明显高于对照组（48.59%比16.34%，n P<0.01），促凋亡蛋白Bax和caspase-3以及促炎因子IL-1β、IL-6、TNF-α的蛋白和mRNA表达水平均高于对照组（均n P<0.01），而抗凋亡蛋白Bcl-2和抗炎因子IL-10的蛋白和mRNA表达水平均低于对照组（n P<0.05，n P<0.01）。n 结论:胺碘酮可导致HUVEC损伤，这种损伤作用随胺碘酮浓度升高和作用时间延长而增强；胺碘酮可能通过诱导细胞凋亡、炎症反应及氧化应激导致HUVEC损伤。“,”Objective:To explore the injury effect and its possible mechanism of amiodarone on human umbilical vein endothelial cells (HUVECs).Methods:After 3 generations of cultivation, the HUVECs were seeded in 96-well plates and incubated with amiodarone (0, 10, 20, 30, and 60 μmol/L) for 24 hours. The cell viability was detected using cell counting kit 8 (CCK-8) assay and the relative viability of cells incubated with different concentrations of amiodarone were calculated by taking the cell viability of the 0 μmol/L group as 100%. The concentration of amiodarone at which cell viability was reduced to 70% was selected for subsequent experiments. The effect of amiodarone of this concentration on the activity of HUVECs after action for different time (6, 12, 24, 36, and 48 hours) was detected using the CCK-8 assay. HUVECs cultured with amiodarone of this concentration were set as the experimental groups and those without amiodarone were set as the control group. Apoptosis rate of HUVECs was detected by Annexin V-FITC/P flow cytometry; the protein and mRNA expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), Caspase-3, interleukin 10 (IL-10), IL-1β, IL-6, and tumor necrosis factor alpha (TNF-α) were detected using western blotting and real-time fluorescence quantitative polymerase chain reaction, respectively; the reactive oxygen species (ROS) was detected by DCFH-DA fluorescence probe assay; the superoxide dismutase (SOD) activity was detected by water-soluble tetrazolium-1 assay; the reduced glutathione (GSH) content was detected by microplate assay.Results:The viabilities of HUVECs incubated with amiodarone at concentration of 10, 20, 30, and 60 μmol/L for 24 hours were (88.82±2.64)%, (74.96±1.75)%, (64.95±2.10)%, and (18.57±0.65)%, respectively; differences were all significant (all n P<0.01) between each experiment group and control group, as well as between each experiment group. Amiodarone at a concentration of 30 μmol/L was used for subsequent experiments. After incubating with 30 μmol/L amiodarone for 6, 12, 24, 36, and 48 hours, the viabilities of HUVECs were (90.19±1.88)%, (82.81±2.51)%, (75.33±1.37)%, (65.76±1.85)%, and (47.01±3.29)%, respectively; differences were all significant (alln P<0.01) between each experiment group and control group, as well as between each experiment group. Compared with the control group, the apoptosis rate of cells in the experimental group was significantly higher (48.59%n vs. 16.34%, n P<0.01), the protein and mRNA expression levels of pro-apoptotic proteins Bax and caspase-3, and pro-inflammatory factors IL-1β, IL-6, and TNF-α were higher (alln P<0.01), whereas the protein and mRNA expression levels of anti-apoptotic protein Bcl-2 and anti-inflammatory factor IL-10 were lower (n P<0.05,n P<0.01).n Conclusions:Amiodarone can cause HUVECs injury, which would be enhanced with the increase of concentration and action time of amiodarone. Amiodarone may cause HUVECs injury by inducing apoptosis, inflammatory response, and oxidative stress.