F2-isoprostanes can mediate bleomycin-induced lung fibrosis

Abstract

F₂-isoprostanes (F₂-IsoPs) have been considered markers of oxidative stress in various pulmonary diseases, but little is known about their possible role in pulmonary fibrosis. In this study, we have investigated the potential key role of F₂-IsoPs as markers and mediators of bleomycin (BLM)-induced pulmonary fibrosis in rats. During the in vivo study, plasma F₂-IsoPs showed a peak at 7 days and remained elevated for the entire experimental period. Lung F₂-IsoP content nearly tripled 7 days following the intratracheal instillation of BLM, and by 28 days, the value increased about fivefold compared to the controls. Collagen deposition correlated with F₂-IsoP content in the lung. Furthermore, from day 21 onwards, lung sections from BLM-treated animals showed α-smooth muscle actin (α-SMA) positive cells, which were mostly evident at 28 days. In vitro studies performed in rat lung fibroblasts (RLF) demonstrated that either BLM or F₂-IsoPs stimulated both cell proliferation and collagen synthesis. Moreover, RLF treated with F₂-IsoPs showed a significant increase of α-SMA expression compared to control, indicating that F₂-IsoPs can readily activate fibroblasts to myofibroblasts. Our data demonstrated that F₂-IsoPs can be mediators of key events for the onset and development of lung fibrosis, such as cell proliferation, collagen synthesis and fibroblast activation. Immunocytochemistry analysis, inhibition and binding studies demonstrated the presence of the thromboxane A₂ receptor (TP receptor) on lung fibroblasts and suggested that the observed effects may be elicited through the binding to this receptor. Our data added a new perspective on the role of F₂-IsoPs in lung fibrosis by providing evidence of a profibrotic role for these mediators in the pathogenesis of pulmonary fibrosis.

Keywords: Bleomycin; F(2)-isoprostanes; Lung fibroblast; Lung fibrosis; Myofibroblasts; Oxidative stress; TP receptor; α-SMA.