Reactive oxygen species provoke secretion of mucin and activate phospholipase c in airway epithelial cells in vitro via a mechanism dependent on nitric oxide. Wright, David T., Bernard M. Fischer, Chengming Li, Lori G. Rochelle, Nancy J. Akley, and Kenneth B. Adler. Department of Toxicology, Department of Anatomy, Physiological Sciences, and Radiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina
APStracts 3:0120L, 1996.
Reactive oxygen species (ROS) have been implicated in the pathogenesis of a wide variety of respiratory diseases. We investigated mechanisms of ROS - induced mucin secretion by guinea pig tracheal epithelial (GPTE) cells in primary culture, and ROS - induced activation of the second messenger - producing enzyme, phospholipase C (PLC), in GPTE cells and in a virally-transformed cell line (BEAS-2B) derived from human bronchial epithelium. Mucin secretion was measured by a monoclonal antibody - based ELISA, and PLC activation was assessed by anion exchange chromatography. ROS generated enzymatically by xanthine oxidase in the presence of purine (P+XO; 500 [mu]M + 20 mU/ml) enhanced release of mucin by GPTE cells and activated PLC in GPTE and BEAS cells. Hypersecretion of mucin and activation of PLC in response to P+XO appeared to occur via an intracellular pathway(s) dependent on endogenously produced nitric oxide (NO) and possibly intracellularly generated oxidants. Both responses could be blocked or attenuated by preincubation of the cells with NG-monomethyl-L -arginine (L-NMA), an inhibitor of the enzyme nitric oxide synthase (NOS), or with dimethylthiourea (DMTU) a compound that can react with a variety of intracellular oxidant species. Reactive nitrogen species (RNS) generated chemically also stimulated secretion of mucin and activated PLC via a mechanism dependent (at least in part) on intracellular oxidant-mediated process(es). The results suggest that intracellularly - generated radical species of nitrogen and oxygen may be important modulators of the response of airway epithelial cells to external oxidant stress.

Received 5 October 1995; accepted in final form 10 July 1996.
APS Manuscript Number L295-5.
Article publication pending Am. J. Physiol. (Lung Cell. Mol.
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 4 August 1996