邻苯二甲酸二(2-乙基己基)酯致人脐静脉内皮细胞氧化损伤及白藜芦醇的修复作用  

Oxidative damage to human umbilical vein endothelial cells caused by di(2-ethylhexyl)phthalate and the repairing effect of resveratrol

作　　者：王子璇 刘伟[1] 蔺博涵 李涛[1] 杨琴[2] 虞绮雯 朱伟[1] 孙晓春[1] WANG Zixuan;LIU Wei;LIN Bohan;LI Tao;YANG Qin;YU Qiwen;ZHU Wei;SUN Xiaochun(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013;Department of Women′s Health,The Fourth Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212000,China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学第四附属医院妇女保健科,江苏镇江212000

出　　处：《江苏大学学报（医学版）》2024年第1期37-43,共7页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81972313)。

摘　　要：目的:探究邻苯二甲酸二(2-乙基己基)酯(di-2-ethylhexyl phthalate,DEHP)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)的氧化损伤,以及白藜芦醇对该损伤的修复作用。方法:通过CCK8实验筛选DEHP的损伤浓度和白藜芦醇的修复浓度;化学分光光度比色法检测HUVEC丙二醛和超氧化物歧化酶(SOD)的不同表达情况;活性氧荧光探针检测不同处理组HUVEC内活性氧水平;JC-1试剂盒检测不同组HUVEC线粒体膜电位;CCK8实验、Transwell实验、划痕愈合实验观察HUVEC增殖、迁移能力;细胞流式实验、蛋白质印迹实验观察HUVEC的凋亡情况和凋亡相关蛋白的表达。结果:80μmol/L DEHP作用HUVEC 24 h后,细胞增殖能力明显降低(P<0.01),40μmol/L白藜芦醇对细胞无明显毒性;损伤组细胞的丙二醛水平和SOD活性较对照组均明显上升(P均<0.01),修复组丙二醛水平和SOD活性较损伤组均明显下降(P均<0.05);损伤组细胞的活性氧表达较对照组明显上升(P<0.01),修复组较损伤组明显下降(P<0.01);损伤组细胞的线粒体膜电位较对照组明显下降(P<0.01),修复组较损伤组明显上升(P<0.01);损伤组细胞的增殖、迁移、成管能力较对照组明显下降(P均<0.05),修复组较损伤组明显上升(P均<0.05);损伤组细胞的凋亡水平较对照组明显上升(P<0.05),修复组较损伤组明显下降(P<0.05)。结论:DEHP能够通过氧化应激途径抑制HUVEC的增殖、迁移和成管能力,并诱导其凋亡,而白藜芦醇可以有效修复该种损伤。Objective:To investigate the oxidative damage of human umbilical vein endothelial cells HUVEC induced by di-2-ethylhexyl phthalate(di-2-ethylhexyl phthalate,DEHP),and the repairing effect of resveratrol on this damage.Methods:The ranges of damage concentration of DEHP and the repair concentration of resveratrol were screened by CCK8 experiment.The different levels of malondialdehyde(MDA)and superoxide dismutase(SOD)activity in HUVEC were detected by chemical spectrophotometry.The content of reactive oxygen species(ROS)in HUVEC was detected by fluorescent probe of reactive oxygen species.The mitochondrial membrane potential(MMP)of different HUVEC groups was detected by JC-1 kit.CCK8 experiment,Transwell experiment and scratch healing experiment were applied to observe the changes of various cell capacities of HUVEC.The apoptosis of HUVEC were detected by cell flow cytometry experiment and Western blotting.Results:After treated with 80μmol/L DEHP for 24 hours,the cell proliferation ability was significantly reduced(P<0.01),and 40μmol/L resveratrol had no obvious toxicity to the cells;The MDA level and SOD activity of the cells in the injury group were significantly higher than those in the control group(all P<0.01),compared with the damage group,the repair group were significantly decreased(all P<0.05);the expression of ROS in the damage group was significantly higher than that in the control group(P<0.01),the repair group was significantly lower than the damage group(P<0.01);the MMP of the damage group was significantly lower than that of the control group(P<0.01),and the repair group was significantly higher than the damage group(P<0.01);Compared with the control group,the proliferation,migration,and tube-forming abilities of the cells were significantly decreased(all P<0.05),and the repair group was significantly increased compared with the damage group(all P<0.05);the apoptosis level of the damage group was significantly higher than that of the control group(P<0.05),the repair group was significantly lower th

关 键 词：人脐静脉内皮细胞 邻苯二甲酸二(2-乙基己基)酯 白藜芦醇 氧化应激 凋亡 氧化损伤修复 

分 类 号：R114[医药卫生—卫生毒理学] R285.5[医药卫生—公共卫生与预防医学]