Mechanistic insights into severe pulmonary inflammation caused by silica stimulation: The role of macrophage pyroptosis

Abstract

Respirable silica dust is a common hazard faced by occupational workers and prolonged exposure to this dust can lead to pulmonary inflammation, fibrosis and, in severe cases, silicosis. However, the underlying mechanism by which silica exposure causes these physical disorders is not yet understood. In this study, we aimed to shed light on this mechanism by establishing in vitro and in vivo silica exposure models from the perspective of macrophages. Our results showed that compared to the control group, silica exposure resulted in an upregulation of the pulmonary expression of P2X7 and Pannexin-1, but this effect was suppressed by treatment with MCC950, a specific inhibitor of NLRP3. Our in vitro studies showed that silica exposure induced mitochondrial depolarization in macrophages, which led to a reduction of intracellular ATP and an influx of Ca2+. Furthermore, we found that creating an extracellular high potassium environment by adding KCl to the macrophage medium inhibited the expression of pyroptotic biomarkers and pro-inflammatory cytokines such as NLRP3 and IL-1β. Treatment with BBG, a P2X7 antagonist, also effectively inhibited the expression of P2X7, NLRP3, and IL-1β. On the other hand, treatment with FCF, a Pannexin-1 inhibitor, suppressed the expression of Pannexin-1 but had no effect on the expression of pyroptotic biomarkers such as P2X7, NLRP3, and IL-1β. In conclusion, our findings suggest that silica exposure triggers the opening of P2X7 ion channels, resulting in intracellular K+ efflux, extracellular Ca2+ influx, and the assembly of the NLRP3 inflammasome, ultimately leading to macrophage pyroptosis and pulmonary inflammation.