Objective To investigate the effects of zinc oxide nanoparticles(ZnO-NPs)on neutrophil hypoxia and pyroptosis through nucleotide binding of oligomeric domain-like receptor protein 3(NLRP3)inflammasome,and to analyze the role of pyroptosis on respiratory tract inflammotion induced by ZnO-NPs.Methods In October 2022,primary cultured neutrophils were obtained from the abdominal aortic blood of SPF adult healthy SD rats.The neutrophils were treated with ZnO-NPs solution(0,5,10,20 μg/ml)at different concentrations,and hypoxia group(5％O2)was set up.Hypoxia and reactive oxygen species(ROS)levels were detected by flow cytometry,and the expression levels of NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),cleaved Caspase-1 were measured by Western blot.The activity of lactic dehydrogenase(LDH)in the cell supernatant was measured by coloration,and the content of interleukin-1 beta(IL-1β)in cell culture supernatant was detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the control group,hypoxia and ROS levels of neutrophils in hypoxia group and ZnO-NPs groups were significantly increased(P＜0.05),and NLRP3,ASC,cleaved Caspase-1 protein expression levels,LDH activity and IL-1β content were significantly increased(P＜0.05).Compared with hypoxia group,hypoxia and ROS levels of neutrophils in 5 µg/ml and 10 µg/ml ZnO-NPs groups were significantly decreased(P＜0.05),NLRP3,ASC,cleaved Caspase-1 protein expression levels,LDH activity,and IL-1β content were decreased significantly(P＜0.05).There were no significant differences in hypoxia,ROS levels,ASC,cleaved Caspase-1 protein expression levels,LDH activity,and IL-1β content between the 20 µg/ml ZnO-NPs group and the hypoxia group(P＞0.05).Conclusion ZnO-NPs treatment may activate the NLRP3 inflammasome to induce pyroptosis of neutrophils which may be related to ROS and hypoxia.
关键词
中性粒细胞/纳米氧化锌/炎症小体/核苷酸结合寡聚化结构域样受体蛋白3/缺氧/焦亡
Key words
Neutrophils/Zinc oxide nanoparticles/Inflammasome/Nucleotide binding of oligomeric domain-like receptor protein 3,NLRP3/Hypoxia/Pyroptosis
维普
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