氧化铟锡纳米颗粒通过氧化应激激活NF-κB/Nrf2通路诱导大鼠肺泡蛋白沉积症  

Nano-ITO induce pulmonary alveolar proteinosis through oxidative stress and activation of NF-κB/Nrf2 signaling pathway

作　　者：林殷乔 张意[2] 陈晓阳 李纬康 牛钰靖 王雪妃 刘楠 李盖 Lin Yinqiao;Zhang Yi;Chen Xiaoyang;Li Weikang;Niu Yujing;Wang Xuefei;Liu Nan;Li Gai(Institute of Public Health,North China University of Science and Technology,Hebei Key Laboratory of Occupational Health and Safety for Coal Industry,Tangshan 063210,China;Quality Control Office,National Institute of Occupational Health and Poison Control,Chinese Center for Disease Control and Prevention,Beijing 100050,China;Medical Imaging Department,North China University of Science and Technology Affiliated Hospital,Tangshan 063000,China)

机构地区：[1]华北理工大学公共卫生学院研究院,河北省煤矿卫生与安全重点实验室,唐山063210 [2]中国疾病预防控制中心职业卫生与中毒控制所质量控制办公室,北京100050 [3]华北理工大学附属医院医学影像科,唐山063000

出　　处：《中华劳动卫生职业病杂志》2025年第2期81-90,共10页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：唐山市科技计划项目(24130225C);河北省中央引导地方科技专项项目(246Z7734G);河北省医学科学研究课题计划资助(20250180);华北理工大学大学生创新创业计划(S202410081007)。

摘　　要：目的建立氧化铟锡纳米颗粒(Nano-ITO)诱导大鼠肺泡蛋白沉积症(PAP)模型,探讨Nano-ITO通过核因子κB(NF-κB)/核转录因子E2相关因子2(Nrf2)通路激活介导大鼠肺PAP的发生规律。方法于2019年10月,随机将120只SD大鼠分为3、7、14、28、56、84 d Nano-ITO染毒组和相应时间点对照组,每组10只;染毒组用6 mg/kg·bw Nano-ITO行非暴露式气管注入染毒,每周2次,进行时间效应研究,分析支气管肺泡灌洗液(BALF)中炎症因子和氧化应激指标水平,苏木精-伊红(HE)、过碘酸-雪夫(PAS)、马松(Masson)、油红O染色法观察肺组织病理形态的改变,透射电镜观察肺组织细胞超微结构,酶联免疫吸附试验(ELISA)、蛋白质免疫印迹(Western blot)法和实时荧光定量聚合酶链反应(RT-PCR)法检测核因子κB p65(NF-κB p65)、核因子κB抑制蛋白α(IκB-α)、核因子κB抑制物激酶(IKK-β)、Nrf2、还原型辅酶Ⅱ醌氧化还原酶1(NQO1)、血红素加氧酶1(HO-1)的定位及表达。两组间比较用独立样本t检验,多组间比较采用单因素方差分析。结果气管内灌注Nano-ITO可诱发大鼠肺组织发生急性炎症、肉芽肿(结节)形成和肺泡蛋白沉积。ELISA分析显示,与相应时间点对照组比较,Nano-ITO染毒组大鼠BALF中超氧化物歧化酶(SOD)、总抗氧化能力(T-AOC)、丙二醛(MDA)、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子α(TNF-α)、IL-10和总蛋白(TP)、乳酸脱氢酶(LDH)的效应指标水平均升高(P<0.05)。大鼠肺组织中Nrf2、NF-κB p65蛋白表达上调,IKK-β的蛋白表达增高(P<0.01)。Nrf2及其下游蛋白NQO1和HO-1在Nano-ITO诱导大鼠PAP中处于高表达状态。结论Nano-ITO暴露可诱导大鼠肺组织发生炎症/氧化应激反应,激活NF-κB/Nrf2信号通路,导致大鼠肺组织发生PAP。ObjectiveTo investigate the role of the nuclear factor-kappa B(NF-κB)/nuclear factor E2 related factor 2(Nrf2)pathway in the occurrence of lung tissue in the pulmonary alveolar proteinosis(PAP)model of rats induced by indium tin oxide nanoprticles(Nano-ITO).MethodsIn October 2019,120 SD rats were divided into 3,7,14,28,56,and 84 day Nano ITO exposure groups and corresponding time point control groups,with 10 rats in each group;the exposure group was treated with 6 mg/kg·bw Nano-ITO via non exposed tracheal injection,twice a week.Time-course studies were performed to examine the pulmonary toxicity induced by Nano-ITO.At the end of the experiment,cytokines levels and oxidative stress were analyzed in the bronchoalveolar lavaged fluid(BALF).Rat lung tissues were also harvested for staining with HE,PAS,Masson,and Oil Red O.Ultrastructure of lung tissue cells was observed by transmission electron microscope.The localization and expression of NF-κB p65,IκB-α,IKK-β,Nrf2,NQO1,HO-1 were observed by immunohistochemistry,Western blot and real-time fluorescent quantitative PCR.The comparison between the two groups was analyzed by independent sample T test,and the comparison between the multiple groups was analyzed by one-way ANOVA.ResultsNano-ITO intratracheal instillation caused pulmonary toxicity by inducing acute inflammation,granuloma(nodule)formation,and alveolar proteinosis.ELISA analysis showed that,compared with the corresponding time points control groups,the levels of superoxide dismutase(SOD),total antioxidant capacity(T-AOC),malondialdehyde(MDA),interleukin(IL)-1β,IL-6,tumor necrosis factor alpha(TNF-α),IL-10,total protein(TP),and lactate dehydrogenase(LDH)in BALF of rats exposed to Nano ITO were all increased(P<0.05);The protein expression of Nrf2 and NF-κB p65 was upregulated in rat lung tissue,while the protein expression of KK-βwas increased(P<0.01).Nrf2 and its downstream proteins NQO1 and HO-1 were highly expressed in Nano-ITO-induced PAP rat.ConclusionNF-κB/Nrf2 signal pathway is involved in the

关 键 词：金属纳米粒子 大鼠 肺泡蛋白沉积症 纳米氧化铟锡 氧化应激 炎症反应 NF-κB/Nrf2通路 

分 类 号：R28[医药卫生—中药学]