Role of superoxide and nitric oxide in platelet-activating factor-induced acute lung injury, hypotension, and mortality in rats.
OBJECTIVE
To investigate the role of superoxide and nitric oxide in platelet-activating factor-induced acute lung injury, hypotension, and mortality.


METHODS
Prospective, randomized, controlled, experimental study.


METHODS
University research laboratory.


METHODS
Anesthetized male Wistar rats (180 to 220 g) were studied.


METHODS
In the first set of experiments, animals were divided into three groups. Group 1 received platelet-activating factor (2 microg/kg i.v.). Group 2 received recombinant human superoxide dismutase (50,000 U/kg i.v.) 30 mins before platelet-activating factor injection. Group 3 received vehicle agents. In the second set of experiments, animals were divided into six groups that received N(G)-nitro-L-arginine (L-NNA), a selective inhibitor of nitric oxide synthesis, or L-arginine, the physiologic precursor of nitric oxide synthesis: a) vehicles (i.v.); b) vehicle plus L-arginine (100 mg/kg i.v.); c) vehicle plus L-NNA (10 mg/kg i.v.); d) vehicle plus platelet-activating factor (2 microg/kg i.v.); e) L-arginine plus platelet-activating factor; and f) L-NNA plus platelet-activating factor. The first intravenous administration was given 5 mins before the second intravenous injection for each group.


RESULTS
In the first set of experiments, vascular labeling with Monastral blue B demonstrated diffuse microvascular injury in the alveolar capillary beds 2 hrs after platelet-activating factor challenge. Thiobarbituric acid-reactive substances in the lung significantly increased at 2 hrs after platelet-activating factor injection. Platelet-activating factor treatment also resulted in an increased concentration of total protein, albumin, and Evans blue dye in bronchoalveolar lavage fluid at 2 hrs after administration, suggesting platelet-activating factor induction of increased alveolar permeability. The platelet-activating factor-induced alveolar microvascular injury, lipid peroxidation, and increased alveolar permeability were inhibited by pretreatment with recombinant human superoxide dismutase. Although L-NNA alone did not affect alveolar permeability in the second set of experiments, L-NNA treatment before platelet-activating factor challenge significantly aggravated platelet-activating factor-induced increased alveolar permeability 2 hrs after platelet-activating factor challenge. Platelet-activating factor also produced a rapid decrease in blood pressure that was not ameliorated by treatment with L-NNA. However, L-NNA pretreatment was associated with a significant increase in platelet-activating factor-caused mortality within 6 hrs. All rats survived with L-arginine treatment before platelet-activating factor challenge. L-NNA treatment decreased nitrate/nitrite concentration, an index of total nitric oxide production, in plasma.


CONCLUSIONS
These results indicate that superoxide, the derived active oxygen species, and lipid peroxidation are implicated in the pathogenesis of platelet-activating factor-induced acute lung injury. Nitric oxide does not play a major role in platelet-activating factor-induced hypotension. Nitric oxide appears to play a protective role in the acute lung injury and mortality induced by platelet-activating factor.
Yoshikawa
T
T
First Department of Medicine, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Japan.
Takano
H
H
Yamamoto
H
H
Nakahashi
Y
Y
Yamaguchi
T
T
Arimoto
T
T
Kondo
M
M
D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007022:Hypotension / Q000139:chemically induced*; D015227:Lipid Peroxidation; D008168:Lung / Q000187:drug effects* / Q000473:pathology*; D008297:Male; D009026:Mortality; D009569:Nitric Oxide / Q000502:physiology*; D019335:Nitroarginine / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000633:toxicity*; D011650:Pulmonary Alveoli / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D013482:Superoxide Dismutase / Q000502:physiology*
D004791:Enzyme Inhibitors; D010972:Platelet Activating Factor; D019335:Nitroarginine; D009569:Nitric Oxide; D013482:Superoxide Dismutase