Human keratinocyte response to 4,4’-methylene diphenyl diisocyanate-glutathione conjugate exposure

Workplace exposure to diisocyanates like 4,4’-methylene diphenyl diisocyanate can cause occupational asthma (MDI-OA), and the underlying biological pathways are still being researched.Although uncertainty remains, evidence supports the hypothesis that dermal exposure to MDI plays an important role in the development of MDI-OA.Gene expression, proteomics, and informatics tools were utilised to characterise changes in expression of RNA and protein in cultured human HEKa keratinocyte cells following exposure to conjugates of MDI with glutathione (MDI-GSH).RT-qPCR analysis using a panel of 39 candidate primers demonstrated 9 candidate genes upregulated and 30 unchanged.HPLC-MS/MS analysis of HEKa cell lysate identified 18 540 proteins across all samples 60 proteins demonstrate statistically significant differential expression in exposed cells, some of which suggest activation of immune and inflammatory pathways.The results support the hypothesis that dermal exposures have the potential to play an important role in the development of MDI-OA. Furthermore, proteomic and gene expression data suggest multiple immune (adaptive and innate) and inflammatory pathways may be involved in the development of MDI-OA. Workplace exposure to diisocyanates like 4,4’-methylene diphenyl diisocyanate can cause occupational asthma (MDI-OA), and the underlying biological pathways are still being researched. Although uncertainty remains, evidence supports the hypothesis that dermal exposure to MDI plays an important role in the development of MDI-OA. Gene expression, proteomics, and informatics tools were utilised to characterise changes in expression of RNA and protein in cultured human HEKa keratinocyte cells following exposure to conjugates of MDI with glutathione (MDI-GSH). RT-qPCR analysis using a panel of 39 candidate primers demonstrated 9 candidate genes upregulated and 30 unchanged. HPLC-MS/MS analysis of HEKa cell lysate identified 18 540 proteins across all samples 60 proteins demonstrate statistically significant differential expression in exposed cells, some of which suggest activation of immune and inflammatory pathways. The results support the hypothesis that dermal exposures have the potential to play an important role in the development of MDI-OA. Furthermore, proteomic and gene expression data suggest multiple immune (adaptive and innate) and inflammatory pathways may be involved in the development of MDI-OA.

职业场所暴露于二异氰酸酯（diisocyanates）类化合物，如4,4'-二苯基甲烷二异氰酸酯（4,4’-methylene diphenyl diisocyanate，简称MDI），可诱发职业性哮喘（occupational asthma，MDI-OA），其潜在的生物学发病机制仍处于研究阶段。尽管目前仍存在一定研究不确定性，但已有证据支持“经皮暴露于MDI是MDI-OA发生发展的重要诱因”这一假说。本研究通过基因表达（gene expression）分析、蛋白质组学（proteomics）及生物信息学工具，对暴露于MDI与谷胱甘肽结合物（conjugates of MDI with glutathione，MDI-GSH）的体外培养人HEKa角质形成细胞（HEKa keratinocyte cells）中RNA与蛋白质的表达变化进行了表征分析。采用包含39条候选引物（primers）的组合开展RT-qPCR（实时定量聚合酶链反应）分析，结果显示9个候选基因表达上调，30个候选基因表达无显著变化。对HEKa细胞裂解液（cell lysate）进行HPLC-MS/MS（高效液相色谱-串联质谱）分析，在所有样本中共鉴定出18540种蛋白质；其中60种蛋白质在暴露组细胞中呈现统计学显著性差异表达，部分差异蛋白提示免疫与炎症通路被激活。本研究结果支持“经皮暴露可能在MDI-OA的发生发展中发挥重要作用”这一假说。此外，蛋白质组学与基因表达数据提示，多种免疫（适应性免疫与固有免疫）及炎症通路可能参与了MDI-OA的发病进程。
职业场所暴露于二异氰酸酯（diisocyanates）类化合物，如4,4'-二苯基甲烷二异氰酸酯（4,4’-methylene diphenyl diisocyanate，简称MDI），可诱发职业性哮喘（occupational asthma，MDI-OA），其潜在的生物学发病机制仍处于研究阶段。尽管目前仍存在一定研究不确定性，但已有证据支持“经皮暴露于MDI是MDI-OA发生发展的重要诱因”这一假说。本研究通过基因表达（gene expression）分析、蛋白质组学（proteomics）及生物信息学工具，对暴露于MDI与谷胱甘肽结合物（conjugates of MDI with glutathione，MDI-GSH）的体外培养人HEKa角质形成细胞（HEKa keratinocyte cells）中RNA与蛋白质的表达变化进行了表征分析。采用包含39条候选引物（primers）的组合开展RT-qPCR（实时定量聚合酶链反应）分析，结果显示9个候选基因表达上调，30个候选基因表达无显著变化。对HEKa细胞裂解液（cell lysate）进行HPLC-MS/MS（高效液相色谱-串联质谱）分析，在所有样本中共鉴定出18540种蛋白质；其中60种蛋白质在暴露组细胞中呈现统计学显著性差异表达，部分差异蛋白提示免疫与炎症通路被激活。本研究结果支持“经皮暴露可能在MDI-OA的发生发展中发挥重要作用”这一假说。此外，蛋白质组学与基因表达数据提示，多种免疫（适应性免疫与固有免疫）及炎症通路可能参与了MDI-OA的发病进程。