PM_(2.5)有机提取物经由铁死亡诱导人支气管上皮细胞损伤的研究  

A study on bronchial epithelial cell injury induced by PM_(2.5)organic extracts through ferroptosis

作　　者：王盟盟 龙玥含 陈圆圆 顾雯[1] 王超[1] 石莹[1] 唐宋 段链[1] WANG Meng-meng;LONG Yue-han;CHEN Yuan-yuan;GU Wen;WANG Chao;SHI Ying;TANG Song;DUAN Lian(China CDC Key Laboratory of Environment and Population Health/National Institute of Environmental Health,Chinese Center for Disease Control and Prevention,Beijing 100021,China;Center for Global Health,Nanjing Medical University)

机构地区：[1]中国疾病预防控制中心环境与人群健康重点实验室/中国疾病预防控制中心环境与健康相关产品安全所,北京100021 [2]南京医科大学全球健康中心

出　　处：《环境卫生学杂志》2024年第4期303-311,361,共10页JOURNAL OF ENVIRONMENTAL HYGIENE

基　　金：首都医科大学环境毒理学北京重点实验室开放研究课题(2022hjd102)。

摘　　要：目的探讨细颗粒物(PM_(2.5))有机提取物能否诱导人支气管上皮细胞铁死亡。方法通过索氏提取法提取PM_(2.5)中有机物作为受试物,使用BEAS-2B细胞,以0.1%DMSO溶液作为溶剂对照,染毒于不同剂量(2.5、5、10和20μg/mL)的PM_(2.5)有机提取物构建细胞染毒模型;使用铁死亡抑制剂(Ferrostatin-1,Fer-1)构建铁死亡干预模型。通过吸光度法检测PM_(2.5)有机提取物染毒后BEAS-2B细胞存活率、丙二醛(malondialdehyde,MDA)和谷胱甘肽(glutathione,GSH)浓度;通过荧光法检测细胞内Fe2+含量、活性氧(reactive oxygen species,ROS)含量和脂质过氧化物(lipid peroxidation,LPO)含量;通过qRT-PCR方法检测铁死亡相关基因(如GPX4、SLC7A11、ACSL4、FTL和TFRC等)表达。结果与对照组相比,PM_(2.5)有机提取物染毒24 h后,当染毒剂量为10μg/mL时,细胞内Fe2+含量、ROS含量、LPO含量和MDA浓度均出现显著上升;而GSH浓度出现显著下降;qRT-PCR结果显示,细胞暴露于20μg/mL PM_(2.5)有机提取物时,细胞内铁死亡相关基因GPX4显著下调,SLC7A11、ACSL4、FTL和TFRC出现显著性上调。干预实验中,使用铁死亡特异性抑制剂Fer-1干预后上述变化得到明显改善。结论PM_(2.5)有机提取物可能通过诱导细胞铁死亡导致呼吸系统细胞损伤,进而对肺组织产生潜在健康影响。Objective This study is intended to investigate whether PM_(2.5)organic extracts can induce ferroptosis in human bronchial epithelial cells.Methods PM_(2.5)organic extracts,obtained through the Soxhlet extraction method,were used as the test poisonous substance.BEAS-2B cells were exposed to different doses of PM_(2.5)organic extracts(2.5,5,10,and 20μg/mL)to establish a cell toxicological model with 0.1%DMSO solution as the control solvent,and the ferroptosis inhibitor(Ferrostatin-1,Fer-1)was used to establish an intervention model.The cell viability of BEAS-2B cells after being exposed to PM_(2.5)organic extracts was evaluated using a spectrophotometric method.The intracellular level of Fe 2+,reactive oxygen species(ROS),lipid peroxidation(LPO),as well as malondialdehyde(MDA)and glutathione(GSH)were measured using fluorescence method.The expression of ferroptosis-related genes(such as GPX4,SLC7A11,ACSL4,FTL,and TFRC)were detected using qRT-PCR.Results Compared to the control group,the concentrations of Fe 2+,ROS,LPO and MDA in BEAS-2B cells exposed to 10μg/mL PM_(2.5)organic extracts for 24 h were significantly increased.Howerver,the concentration of GSH decreased with the exposure dose.qRT-PCR result showed that cells with 20μg/mL of PM_(2.5)organic extracts significantly downregulated the ferroptosis-related gene GPX4,while upregulated SLC7A11,ACSL4,FTL,and TFRC.In the intervention experiment,there is convincing evidence suggesting that the use of the iron death-specific inhibitor Fer-1 was able to significantly improve the injury caused by PM_(2.5)organic extracts.Conclusion The organic extracts of PM_(2.5)may potentially induce cellular ferroptosis,resuting in respiratory system cell damage,thereby exerting potential health impacts on lung tissues.To investigate whether PM_(2.5)organic extract can induce ferroptosis in human bronchial epithelial cells.Methods PM_(2.5)organic extract was obtained using the Soxhlet extraction method and used as the poisonous substance.BEAS-2B cells were exposed to different

关 键 词：细颗粒物(PM_(2.5)) 有机提取物 铁死亡 人支气管上皮细胞 肺损伤 

分 类 号：R122[医药卫生—环境卫生学]