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N-乙酰半胱氨酸对PM2.5致支气管上皮细胞损伤的保护作用体外实验

郭淼 闫鹏 韩国鑫 朱明 金珊 肖坤 解立新

郭淼, 闫鹏, 韩国鑫, 朱明, 金珊, 肖坤, 解立新. N-乙酰半胱氨酸对PM2.5致支气管上皮细胞损伤的保护作用体外实验[J]. 解放军医学院学报, 2021, 42(1): 99-103. doi: 10.3969/j.issn.2095-5227.2021.01.021
引用本文: 郭淼, 闫鹏, 韩国鑫, 朱明, 金珊, 肖坤, 解立新. N-乙酰半胱氨酸对PM2.5致支气管上皮细胞损伤的保护作用体外实验[J]. 解放军医学院学报, 2021, 42(1): 99-103. doi: 10.3969/j.issn.2095-5227.2021.01.021
GUO Miao, YAN Peng, HAN Guoxin, ZHU Ming, JIN Shan, XIAO Kun, XIE Lixin. Protective effect of N-acetylcysteine on PM2.5-induced injury of bronchial epithelial cells in vitro[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL, 2021, 42(1): 99-103. doi: 10.3969/j.issn.2095-5227.2021.01.021
Citation: GUO Miao, YAN Peng, HAN Guoxin, ZHU Ming, JIN Shan, XIAO Kun, XIE Lixin. Protective effect of N-acetylcysteine on PM2.5-induced injury of bronchial epithelial cells in vitro[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL, 2021, 42(1): 99-103. doi: 10.3969/j.issn.2095-5227.2021.01.021

N-乙酰半胱氨酸对PM2.5致支气管上皮细胞损伤的保护作用体外实验

doi: 10.3969/j.issn.2095-5227.2021.01.021
详细信息
    作者简介:

    郭淼,男,本科,主治医师。研究方向:气道炎症和损伤,危重症诊治。Email: guom306em@163.com

    通讯作者:

    解立新,男,博士,主任医师,解放军总医院呼吸与危重症医学部主任。Email: xielx301@126.com

  • 中图分类号: R 965

Protective effect of N-acetylcysteine on PM2.5-induced injury of bronchial epithelial cells in vitro

More Information
  • 摘要:   背景  大气污染物中的PM2.5可以引起气道上皮细胞炎症反应,N-乙酰半胱氨酸(N-acetyl-L-cysteine,NAC)可以有效减轻这种炎症反应,但相关的自噬机制研究较少。通过对自噬机制的研究,可为抑制PM2.5诱导的气道炎症反应提供新思路。  目的  探讨N-乙酰半胱氨酸对PM2.5引起的支气管上皮细胞损伤的保护机制。  方法  将人支气管上皮细胞(BEAS-2B)按照不同暴露因素设为控制对照组、PM2.5组、PM2.5+NAC组。对照组使用普通培养基处理16 h;PM2.5组用普通培养基预处理4 h后再用含有PM2.5的培养基处理12 h;PM2.5+NAC组用含有NAC的培养基预处理4 h再用含有PM2.5的培养基处理12 h。流式细胞仪分析细胞凋亡情况,使用荧光共聚焦显微镜观察LC3荧光量,Western blot法观察LC3Ⅱ/LC3Ⅰ及P62的情况,分析三组细胞发生自噬的情况。  结果  与对照组和PM2.5+NAC组比较,PM2.5组细胞凋亡明显增加;而PM2.5+NAC组由PM2.5(100 μg/mL)引起的BEAS-2B细胞凋亡明显被抑制。免疫荧光结果显示,与对照组比较,PM2.5组LC3荧光量明显增加;与PM2.5组比较,PM2.5+NAC组LC3荧光量明显减少。Western blot结果显示,与对照组比较,PM2.5组LC3Ⅱ/LC3Ⅰ明显增加,P62明显减少;与PM2.5组比较,PM2.5+NAC组LC3Ⅱ/LC3Ⅰ明显减少,P62明显增加。  结论  NAC可以抑制PM2.5介导的人支气管上皮细胞的凋亡和自噬,从而减少细胞的损伤。

     

  • 图  1  流式细胞仪测定细胞凋亡及定量分析 (aP<0.05,vs PM2.5+NAC;bP<0.05,vs PM2.5

    Figure  1.  Flow cytometry results of apoptosis and quantification analysis (aP<0.05, vs PM2.5+NAC; bP<0.05, vs PM2.5

    图  2  细胞自噬荧光图

    A:细胞核的DAPI染色(蓝色)、细胞骨架的鬼笔环肽染色(红色)作为背景染色,LC3荧光呈绿色;B:荧光定量分析(aP<0.05,vs PM2.5;bP<0.05,vs PM2.5+NAC)

    Figure  2.  Measurement of autophagy by fluorescent microscopy

    A: DAPI staining of nuclei (blue) and Phalloidin staining of cytoskeleton (red) were set as the background stain, and LC3 fluorescence was green; B: Quantification analysis (aP<0.05, vs PM2.5; bP<0.05, vs PM2.5+NAC)

    图  3  LC3Ⅰ、LC3Ⅱ、P62、GAPDH定量分析(A。aP<0.05,vs PM2.5;bP<0.05,vs PM2.5+NAC)及化学发光结果(B)

    Figure  3.  Quantification analysis (A. aP<0.05, vs PM2.5; bP<0.05, vs PM2.5+NAC) and chemiluminescence results of LC3Ⅰ, LC3Ⅱ, P62 (B)

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出版历程
  • 收稿日期:  2020-09-08
  • 网络出版日期:  2021-02-10
  • 刊出日期:  2021-01-28

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