亚低温对聚苯乙烯微塑料所致血管内皮细胞损伤的影响

Effects of mild hypothermia on vascular endothelial cell injury induced by polystyrene microplastics

目的探讨亚低温在聚苯乙烯微塑料(polystyrene microplastics,PS-MPs)所致血管内皮细胞损伤中的保护作用。方法用含10%胎牛血清的DMEM培养基常规培养人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)。对HUVEC给予PS-MPs暴露和亚低温处理。实验分组为:对照组、PS-MPs处理组、33℃亚低温组、PS-MPs+33℃亚低温处理组。采用CCK-8法检测细胞存活率,流式细胞术检测活性氧(reactive oxygen species,ROS)水平,使用试剂盒检测超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)水平,Real time PCR和Western blot法检测凋亡基因Bax和Bcl-2的mRNA和蛋白表达水平。结果HUVEC细胞暴露于PS-MPs后,细胞ROS、MDA水平和Bax蛋白表达量均显著升高(P<0.05)。与PS-MPs处理组相比,亚低温处理后,ROS、MDA水平以及Bax的mRNA和蛋白表达水平均降低(P<0.05),SOD水平升高(P<0.05),HUVEC凋亡状况减轻。结论PS-MPs可引起血管内皮细胞的氧化应激和细胞凋亡,损伤血管内皮细胞;亚低温可通过抑制氧化损伤和细胞凋亡,发挥保护作用。

Objective To(PS-MPs)on human vascular endothelial cells,and to explore the protective effect of mild hypothermia against the injury of vascular endothelial cells caused by polystyrene microplastics(PS-MPs).Methods Human umbilical vein endothelial cells(HUVECs)were routinely cultured in DMEM medium containing 10%fetal bovine serum.HUVECs were exposed to PS-MPs and treated with mild hypothermia.The experimental groups were as follows:control group,PS-MPs exposure group,33℃mild hypothermia treatment group,and PS-MPs+33℃mild hypothermia treatment group.The cell counting kit-8 assay was used to detect the cell survival rate,and flow cytometry was used to measure reactive oxygen species(ROS)level.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)were measured by corresponding kits.Real-time polymerase chain reaction and Western blot were used to determine mRNA and protein expression levels of apoptotic genes of Bax and Bcl-2.Results The ROS and MDA levels and Bax protein expression of HUVECs were significantly increased after exposure to PS-MPs(P<0.05).Compared with the PS-MPs exposure group,the mild hypothermia treatment groups had significantly decreased the levels of ROS and MDA and the mRNA and protein expression levels of Bax(P<0.05),significantly increased SOD levels(P<0.05),and alleviated apoptosis of HUVECs.Conclusion PS-MPs can cause oxidative stress and apoptosis of vascular endothelial cells,then damage vascular endothelial cells.Mild hypothermia plays a protective role by inhibiting oxidative damage and apoptosis.