研究报告

  • 洪启浩,王国镇,冯董董,刘紫佳,唐铎,薛姗,赵子杰,王谦,姚孟菲,侯潇楠,任阵海,周志祥.北京市冬季空气中PM2.5对THP-1细胞焦亡的影响[J].环境科学学报,2022,42(11):475-481

  • 北京市冬季空气中PM2.5对THP-1细胞焦亡的影响
  • Effects on the pyroptosis of THP-1 cells induced by airborne PM2.5 from Beijing in winter
  • 基金项目:国家自然科学基金(No.42077399)
  • 作者
  • 单位
  • 洪启浩
  • 北京工业大学,环境与生命学部,北京 100124
  • 王国镇
  • 北京工业大学,环境与生命学部,北京 100124;中日友好医院,检验科,北京 100029
  • 冯董董
  • 北京工业大学,环境与生命学部,北京 100124
  • 刘紫佳
  • 北京工业大学,环境与生命学部,北京 100124
  • 唐铎
  • 北京工业大学,环境与生命学部,北京 100124
  • 薛姗
  • 北京工业大学,环境与生命学部,北京 100124
  • 赵子杰
  • 北京工业大学,环境与生命学部,北京 100124
  • 王谦
  • 北京工业大学,环境与生命学部,北京 100124
  • 姚孟菲
  • 北京工业大学,环境与生命学部,北京 100124
  • 侯潇楠
  • 北京工业大学,环境与生命学部,北京 100124
  • 任阵海
  • 北京工业大学,环境与生命学部,北京 100124
  • 周志祥
  • 北京工业大学,环境与生命学部,北京 100124
  • 摘要:为了验证PM2.5进入肺部后,在肺巨噬细胞清除PM2.5组分中是否发生细胞焦亡效应,本研究以THP-1细胞作为人巨噬细胞模型,以不同浓度PM2.5暴露于THP-1细胞,采用CCK-8法检测细胞存活率,试剂盒检测乳酸脱氢酶(LDH)释放水平,荧光显微镜下观察细胞PI染色情况,流式细胞仪检测Annexin V/PI染色,蛋白免疫印迹(Western blotting)检测焦亡相关蛋白NLRP3、ASC、caspase-1、GSDMD的表达,以及ELISA法检测IL-1β、IL-18的分泌等来检测PM2.5暴露诱导THP-1细胞的焦亡发生和炎症因子释放水平.结果显示:THP-1细胞暴露于PM2.5后,细胞存活率的降低与PM2.5暴露浓度的增加呈正相关;THP-1暴露PM2.5 48 h后,细胞胀大并呈气泡状;与空白对照(PBS)组相比,PM2.5暴露组与阳性对照组(1.0 μg·mL-1 LPS+5.0 mmol·L-1 ATP)的细胞上清中LDH水平显著提高;流式细胞仪检测显示Annexin V/PI染色双阳区的细胞比例显著提高;Western blotting结果显示,与对照组相比,PM2.5暴露组炎症小体蛋白NLRP3、ASC、caspase-1表达水平显著提高,而且caspase-1和焦亡执行蛋白GSDMD发生了切割;ELISA结果显示,与对照组相比,PM2.5暴露组IL-1β与IL-18分泌显著提高.研究表明,PM2.5暴露可诱导THP-1细胞炎症性细胞焦亡效应.
  • Abstract:To explore the pyroptosis of macrophage cells induced by PM2.5 exposure, in this study, THP-1 cells were used as human macrophage cell model to investigate PM2.5-induced pyroptosis. THP-1 cells were exposed to different comcentration of PM2.5, the viability of THP-1 cells was detected using a Cell Counting Kit-8 assay. Lactate dehydrogenase (LDH) in the culture medium of THP-1 cells were measured using a colorimetric assay. In the THP-1 cells, the levels of IL-1β and IL-18 were detected using ELISA. The pyroptosis of THP-1 cells was detected using Annexin V/PI double-positive staining. Expression of proteins associated with the nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain-containing protein 3 inflammasome (NLRP3), including NLRP3, apoptosis associated speck-like protein (ASC), stimulated the cleavage of caspase-1 and the pyroptosis executive protein Gasdermin D (GSDMD), were measured by western blotting. PM2.5 significantly inhibited cell viability, induced cell swelling and cell membrane rupture, and increased LDH activity in THP-1 cells. In addition, PM2.5 markedly promoted the production of IL-1β and IL-18 by THP-1 cells. Treatment with PM2.5 significantly increased the protein of NLRP3、ASC、pro-caspase-1, Caspase-1, GSDMD, and cleaved gasdermin D (GSDMD-N) in THP-1 cells. The results from the present study suggest that PM2.5 can induce the pyroptosis in THP-1 cells.

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