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Institute for Environmental Medicine and Department of Biochemistry and Biophysics, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Nitric oxide (· NO) can be produced
within the lung, and recently inhaled nitric oxide has been used as a
therapeutic agent. Peroxynitrite1
(ONOO
), the product of
the nearly diffusion-limited reaction between · NO and
superoxide, may represent the proximal reactive species mediating
· NO injury to pulmonary cells. To investigate the
physiological and pathological reactivities of · NO and
ONOO at the molecular and
cellular levels, bovine pulmonary artery endothelial cells (BPAEC) and
rat type II epithelial cells were exposed to · NO
(0.01-2.5 µM/min for 2 h) generated by spermine-NONOate and
papa-NONOate and to the same fluxes of
ONOO generated by
1,3-morpholinosydnonimine (SIN-1). Exposure to SIN-1 resulted in
cellular injury and death in both cell types. Epithelial cells
displayed a concentration-dependent loss of cellular viability within 8 h of exposure. In contrast, BPAEC loss of cellular viability was
evident after 18 h postexposure. Events preceding cell death in BPAEC
include depolarization of the mitochondrial membrane, evident as early
as 6 h postexposure, loss of cellular redox activity at 16 h, and DNA
fragmentation detected by in situ staining at 18 h after exposure.
Exposure of BPAEC to · NO did not affect the cellular
viability, but type II cells were injured in a manner similar to
ONOO exposure.
· NO-mediated cellular injury within type II cells was
reduced by preincubation with
N-acetylcysteine. The data imply that
the pathological and physiological effects of · NO may be regulated by its reactions with superoxide and reduced thiols.
apoptosis; superoxide; endothelium; type II epithelium
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