Involvement of oxidative stress in crystalline silica-induced cytotoxicity and genotoxicity in rat alveolar macrophages

Abstract

Alveolar macrophages (AMs) occupy a key position in silica-induced pulmonary fibrosis, although the mechanism3 are yet to be elucidated. In the present study we examined the involvement of oxidative stress and reactive oxygen species formation in silica-induced cytotoxicity and genotoxicity in cultured rat AMs. A lucigenin-dependent chemiluminescence test was used to determine superoxide anion (O2/-), and a 2',7'-dichlorofluorescin diacetate fluorescence test was employed to measure the hydrogen peroxide (H2O2) level. The cytotoxic and genotoxic effects caused by silica in AMs were examined by lactate dehydrogenase (LDH) leakage and single-cell gel electrophoresis (comet assay), respectively. The results showed that silica enhanced O2/- and H2O2 formation in AMs. There were clear dose- and time- dependent relationships in silica-induced cytotoxicity and genotoxicity. Furthermore, superoxide dismutase and catalase were able to reduce silica- induced LDH leakage and DNA damage, with concurrent significant inhibition on silica-induced oxidative stress in AMs. These findings provide convincing evidence that oxidative stress mediates the silica-induced cytotoxicity and genotoxicity. The understanding of such a mechanism may provide a scientific basis for the possible application of antioxidants in preventing the hazardous effects of silica. (C) 2000 Academic Press.