纳米氧化锌对大鼠中性粒细胞缺氧和焦亡的影响  

Effect of zinc oxide nanoparticles on hypoxia and pyroptosis of rat neutrophils

作　　者：刘玉梅[1] 肖莎[1] 于德娥[1] 张静 刘云儒[1] 燕贞 Liu Yumei;Xiao Sha;Yu De'e;Zhang Jing;Liu Yunru;Yan Zhen(School of Public Health,Hainan Medical University,Haikou 571199,China)

机构地区：[1]海南医学院公共卫生学院,海口571199

出　　处：《中华劳动卫生职业病杂志》2024年第7期481-486,共6页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：海南省自然科学基金项目(822MS078);国家自然科学基金项目(82160628)。

摘　　要：目的探讨纳米氧化锌(ZnO-NPs)可否通过核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体对中性粒细胞缺氧和焦亡产生影响,分析细胞焦亡在ZnO-NPs诱发呼吸道炎症中的作用。方法于2022年10月,采集SPF级成年健康SD大鼠腹主动脉血,分离并原代培养中性粒细胞,将不同浓度的ZnO-NPs溶液(0、5、10、20μg/ml)作用于中性粒细胞,同时设缺氧(5%O2)组。用流式细胞仪检测缺氧和活性氧(ROS)水平,Western blot法检测细胞中NLRP3、凋亡相关斑点样蛋白(ASC)、活化半胱氨酸蛋白酶-1(cleaved Caspase-1)表达水平。采用比色法检测细胞培养上清液中乳酸脱氢酶(LDH)活力,采用酶联免疫吸附(ELISA)法检测细胞培养上清液中白细胞介素-1β(IL-1β)的含量。结果与对照组比较,缺氧组和各浓度ZnO-NPs组中性粒细胞缺氧和ROS水平均明显升高(P<0.05),细胞NLRP3、ASC、cleaved Caspase-1蛋白表达水平、LDH活力和IL-1β含量均明显升高(P<0.05)。与缺氧组比较,5μg/ml、10μg/ml ZnO-NPs组中性粒细胞缺氧和ROS水平均明显下降(P<0.05),细胞NLRP3、ASC、cleaved Caspase-1蛋白表达水平、LDH活力和IL-1β含量均明显下降(P<0.05)。20μg/ml ZnO-NPs组与缺氧组中性粒细胞缺氧、ROS水平、ASC、cleaved Caspase-1蛋白表达,以及LDH活力和IL-1β含量差异均无统计学意义(P>0.05)。结论ZnO-NPs可激活NLRP3炎症小体从而诱发中性粒细胞焦亡,可能与ROS和缺氧有关。ObjectiveTo 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.MethodsIn 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(0,5,10,20μg/ml)at different concentrations,and hypoxia group(5%O 2)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).ResultsCompared 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-1βcontent were significantly increased(P<0.05).Compared with hypoxia group,hypoxia and ROS levels of neutrophils in 5μg/ml and 10μg/ml ZnO-NPs groups were significantly decreased(P<0.05),NLRP3,ASC,cleaved Caspase-1 protein expression levels,LDH activity,and IL-1βcontent 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-1βcontent between the 20μg/ml ZnO-NPs group and the hypoxia group(P>0.05).ConclusionZnO-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 

分 类 号：R392[医药卫生—免疫学]