ABSTRACT: In the initiation and progression of pulmonary inflammation, macrophages have classically been considered as a crucial cell type. However, evidence for the role of epithelial type II cells in pulmonary inflammation has been accumulating. In the current study, a combined in vivo and in vitro approach has been employed to investigate the mechanisms of quartz-induced proinflammatory activation of lung epithelial cells. In vivo, enhanced expression of the inflammation- and oxidative stress-related genes HO-1 and iNOS was found on the mRNA level in rat lungs after instillation with DQ12 respirable quartz. Activation of the classical NF-kappaB pathway in macrophages and type II pneumocytes was indicated by enhanced immunostaining of phospho-IkappaBalpha in these specific lung cell types. In vitro, the direct, particle-mediated effect on proinflammatory signalling in a rat lung epithelial (RLE) cell line was compared to the indirect, macrophage product-mediated effect. Treatment with quartz particles induced HO-1 and COX-2 mRNA expression in RLE cells in an NF-kappaB independent manner. Supernatant from quartz-treated macrophages rapidly activated the NF-kappaB signalling pathway in RLE cells and markedly induced iNOS mRNA expression up to 2000-fold compared to non-treated control cells. Neutralisation of TNFalpha and IL-1beta in macrophage supernatant did not reduce its ability to elicit NF-kappaB activation of RLE cells. In addition the effect was not modified by depletion or supplementation of intracellular glutathione. The results from the current work suggest that although both oxidative stress and NF-kappaB are likely involved in the inflammatory effects of toxic respirable particles, these phenomena can operate independently on the cellular level. This might have consequences for in vitro particle hazard testing, since by focusing on NF-kappaB signalling one might neglect alternative inflammatory pathways.
van Berlo, D., Knaapen, A. M., van Schooten, F. J., Schins, R. P., & Albrecht, C. (2010). NF-kappaB dependent and independent mechanisms of quartz-induced proinflammatory activation of lung epithelial cells. Particle and Fibre Toxicology, 7(1), 13. https://doi.org/10.1186/1743-8977-7-13