纳米氧化锌诱导血管内皮细胞凋亡及氧化应激

目的探讨纳米氧化锌诱导血管内皮细胞(ECV304)凋亡与活性氧(ROS)的关系。方法采用龙胆紫染色观察细胞吞噬效应;四甲基偶氮噻唑蓝(MTT)法测定细胞生长活性;琼脂糖凝胶电泳检测细胞凋亡;紫外分光光度计测定超氧化物歧化酶(SOD)、丙二醛(MDA)的含量;荧光分光光度计测定ROS。结果 ECV304细胞可吞噬纳米氧化锌,实验组MTT值(0.85±0.04)、(0.56±0.03)、(0.51±0.05)、(0.45±0.044)与对照组(1.21±0.11)比较,差异有统计学意义(P<0.01);实验组ROS(33.14±0.38)、(35.82±0.61)、(54.13±0.24)、(68.81±1.24)均高于对照组(30.12±1.23);实验组SOD(25.72±0.48)、(25.64±0.36)、(25.53±0.36)、(25.25±0.44)均低于对照组(26.39±0.40);实验组MDA含量均高于对照组(0.56±0.01);a-硫辛酸组细胞凋亡率明显降低。结论纳米氧化锌可以抑制ECV304细胞的活性,通过ROS途径诱导细胞凋亡,产生细胞毒性。

Induction of apoptosis in ECV304 cells by zinc oxide nanoparticles

Objective To investigate the apoptosis in ECV304 induced by zinc oxide(ZnO) nanoparticles and the role of reactive oxygen species(ROS) in apoptosis.Methods Cell phagocytic effect was detected using crystal violet staining.Cell inhibition rate was detected by thiazolyl blue tetrazolium bromide(MTT) assay.The rate of DNA damage was detected by DNA ladder gel electrophoresis.The superoxide dismatase(SOD) and malondialdehyde(MDA) were detected by UV spectrophotometer.The ROS was detected by 2’,7’-dichlorofluorescein-diacetate(DCFH-DA) fluorescent probe.Results ZnO nanoparticle entered ECV304 cell through phagocytosis and distributed in cluster.The values of MTT in different dose groups(0.85±0.04,0.56±0.03,0.51±0.05,and 0.45±0.044) were significantly different from that of the control group(P<0.01).The levels of ROS in different dose groups(33.14±0.38,35.82±0.61,54.13±0.24,and 68.81±1.24) were significantly higher than that of the control group(30.12±1.23,P<0.01).The levels of SOD in different dose groups(25.72±0.48,25.64±0.36,25.53±0.36,and 25.25±0.44) were significantly lower than that of the control group(26.39±0.40,P<0.01).The levels of MDA in different dose groups were significantly higher than that of the control group(0.56±0.01,P<0.01).The cell apoptosis of alpha hipoic acid(LA) group was significantly lower than those of treatment groups.Conclusion Nano-ZnO can inhibit ECV304 cell proliferation,induce apoptosis through ROS pathway,and produce cytotoxicity.