Seriani Robson, Junqueira Mara de Souza, de Toledo Alessandra Choqueta, Martins Milton Arruda, Seckler Marcelo, Alencar Adriano Mesquita, Negri Elnara Marcia, Silva Luiz Fernando Ferraz, Mauad Thaís, Saldiva Paulo Hilário Nascimento, Macchione Mariangela
Laboratory of Experimental Air Pollution, Department of Pathology, School of Medicine, University of São Paulo, São Paulo, SP, Brazil.
Central Biotery Laboratory, School of Medicine, University of São Paulo, São Paulo, SP, Brazil.
Environ Toxicol. 2015 Nov;30(11):1297-308. doi: 10.1002/tox.22000. Epub 2014 Apr 29.
Particulate matter from diesel exhaust (DEP) has toxic properties and can activate intracellular signaling pathways and induce metabolic changes. This study was conducted to evaluate the activation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) and to analyze the mucin profile (acid (AB(+) ), neutral (PAS(+) ), or mixed (AB/PAS(+) ) mucus) and vacuolization (V) of tracheal explants after treatment with 50 or 100 μg/mL DEP for 30 or 60 min. Western blot analyses showed small increases in ERK1/2 and JNK phosphorylation after 30 min of 100 μg/mL DEP treatment compared with the control. An increase in JNK phosphorylation was observed after 60 min of treatment with 50 μg/mL DEP compared with the control. We did not observe any change in the level of ERK1/2 phosphorylation after treatment with 50 μg/mL DEP. Other groups of tracheas were subjected to histological sectioning and stained with periodic acid-Schiff (PAS) reagent and Alcian Blue (AB). The stained tissue sections were then subjected to morphometric analysis. The results obtained were compared using ANOVA. Treatment with 50 μg/mL DEP for 30 min or 60 min showed a significant increase (p < 0.001) in the amount of acid mucus, a reduction in neutral mucus, a significant reduction in mixed mucus, and greater vacuolization. Our results suggest that compounds found in DEPs are able to activate acid mucus production and enhance vacuolization and cell signaling pathways, which can lead to airway diseases.
柴油废气中的颗粒物（DEP）具有毒性，可激活细胞内信号通路并诱导代谢变化。本研究旨在评估细胞外信号调节激酶（ERK）和c-Jun氨基末端激酶（JNK）的激活情况，并分析用50或100μg/mL DEP处理30或60分钟后气管外植体的粘蛋白谱（酸性（AB(+)）、中性（PAS(+)）或混合性（AB/PAS(+)）黏液）和空泡化（V）情况。蛋白质免疫印迹分析显示，与对照组相比，100μg/mL DEP处理30分钟后ERK1/2和JNK磷酸化水平略有增加。与对照组相比，50μg/mL DEP处理60分钟后观察到JNK磷酸化增加。在用50μg/mL DEP处理后，我们未观察到ERK1/2磷酸化水平有任何变化。其他组的气管进行组织切片，并用高碘酸-希夫（PAS）试剂和阿尔辛蓝（AB）染色。然后对染色的组织切片进行形态计量分析。使用方差分析对所得结果进行比较。用50μg/mL DEP处理30分钟或60分钟后，酸性黏液量显著增加（p < 0.001），中性黏液减少，混合黏液显著减少，空泡化程度更高。我们的结果表明，DEP中发现的化合物能够激活酸性黏液的产生，增强空泡化和细胞信号通路，这可能导致气道疾病。
