Toxicological effects of air settled particles from the Vitoria Metropolitan Area mediated by oxidative stress, pro-inflammatory mediators and NFΚB pathway

Abstract

Atmospheric pollution is a major environmental and public health risk due to its effect on global air quality and climate. Increase in pollutants concentrations, especially particulate matter (PM), are associated with increased respiratory diseases. The pathophysiology of respiratory diseases involves molecular and cellular mechanisms as inflammatory biomarkers and reactive oxygen species production. Thus, the present study aimed to investigate the in vitro cytotoxic and pro-inflammatory effects of particulate matter (PM) of six monitoring stations (1–6) from the Vitoria Metropolitan Area (VMA), Espirito Santo, Brazil in 2018. The PM was chemically characterized by inductively coupled plasma mass spectrometry. In vitro cytotoxic effects of PM (3.12–200.0 μg/mL) were analyzed in human lung epithelial cells (A549) and macrophage cells (RAW 264.7) by MTT assay (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide). To investigate the pro-inflammatory effects of PM in RAW 264.7 cells, the levels of proinflammatory mediators such as nitric oxide (NO), superoxide anion (O2•-), tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and the activation of nuclear factor kappa B (NF- κB) were measured. The comet assay evaluated genotoxicity. Cell cycle, oxidative stress (DCF and DHE), and apoptosis were analyzed by flow cytometry. Chemical analysis of PM revealed aluminum (Al) and Iron (Fe) as the major chemical elements in all studied monitoring stations. In addition, worrying concentrations of mercury (Hg) were detected in the PM. The in vitro results showed that PM presents a dose-dependent cytotoxic effect in macrophage and pulmonary epithelial cell lines. The PM increased the production of NO, O2•-, and pro-inflammatory cytokines TNF-α and IL-6. PM also promoted alterations in the cell cycle, increased apoptosis frequency, and DNA damage. Moreover, PM increased the expression NF-κB. In addition, a positive correlation between Al and Fe and ROS production was observed. Based on the results obtained during the study period, it was concluded that the sedimented particles from the VMA might have deleterious effects on human health, which was evidenced by the increase in oxidative stress, an increase in pro-inflammatory mediators, and genotoxic effects partially mediated by the NF-κB pathway. These results add aspects to elucidate the molecular mechanisms involved in the effects of sedimented particles in vivo and in vitro.