Am. J. Respir. Cell Mol. Biol., Volume 19, Number 2, August, 1998 226-236

Asbestos Fibers and Interleukin-1 Upregulate the Formation of Reactive Nitrogen Species in Rat Pleural Mesothelial Cells

Nonghoon Choe, Shogo Tanaka, and Elliott Kagan

Department of Pathology, Uniformed Services University of the Health Sciences, F. Edward Hébert School of Medicine, Bethesda, Maryland

Nitric oxide radical (·NO) and peroxynitrite anion (ONOO) have been implicated in lung inflammation and may be important in pleural injury. The present study was undertaken to determine the effects of asbestos exposure and cytokine stimulation on ·NO and ONOO production by rat pleural mesothelial cells. Accordingly, rat parietal pleural mesothelial cells were cultured for 2 to 72 h with or without 50 ng/ml of recombinant interleukin-1 (IL-1) in the presence (1.05 to 8.4 µg/cm²) or absence of crocidolite or chrysotile asbestos fibers. The effects of asbestos were compared with those of carbonyl iron, a nonfibrogenic particulate. Mesothelial cell messenger RNA (mRNA) expression of the inducible form of ·NO synthase (iNOS), assessed with the reverse transcription-polymerase chain reaction (RT-PCR), increased progressively from 2 to 12 h in IL-1-containing cultures. Nitrite (NO₂), the stable oxidation product of ·NO in mesothelial cell conditioned medium, was assayed through the Griess reaction. Both types of asbestos fibers (chrysotile > crocidolite) upregulated the formation of NO₂ in mesothelial cells costimulated with IL-1 in a concentration-dependent and time-dependent fashion. In contrast, carbonyl iron did not upregulate NO₂ formation in IL-1-stimulated cells. Both types of asbestos fibers also induced iNOS protein expression and the formation of nitrotyrosine in mesothelial cells and greatly induced the formation of nitrate (NO₃), a surrogate marker of ONOO formation, in IL-1-stimulated cells. However, the effects of chrysotile were notably greater than those of crocidolite. These findings may have significance for the induction of pleural injury by asbestos fibers.

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