Differential expression of arginase and iNOS in the lung in sepsis.
The primary metabolic fates of L-arginine are conversion to L-citrulline by nitric oxide synthase (NOS) and to L-ornithine by arginase. In the lung, arginine utilization is increased after the inducible form of NOS (iNOS) is expressed during inflammation. The expression of arginase in normal lung and after sepsis, and its potential relationships with iNOS, however, are not known. Since arginase and iNOS share the substrate L-arginine, we tested the hypothesis that lung arginase would be co-induced with iNOS in sepsis and its cellular distribution would be related to that of iNOS in the lung. Lungs from cecal ligation and puncture (CLP) and sham-operated (S) rats were harvested 6 or 16 hours after the procedures. Lung wet-to-dry weight ratio, myeloperoxidase content, and lipid peroxidation products were measured as indices of lung injury. Western blot analyses were performed with polyclonal antibodies against two isoforms of rat arginase (I and II) and iNOS. Additional lungs from CLP and S animals were inflation-fixed for immunohistochemistry using the same antibodies. We found by Western blot that arginase II at 39 kDa was the main isoform present in normal rat lung. The enzyme was distributed diffusely in alveolar and bronchial epithelial cells, endothelial cells, and alveolar macrophages. After CLP, arginase II was almost undetectable in rat lungs at 16 hours. In contrast, in normal lung, the iNOS was not detectable by Western blot or immunohistochemistry. After CLP, strong expression of iNOS was found in similar cell types to arginase II. These data demonstrate loss of constitutive expression of arginase II in rat lung as iNOS is upregulated by the response to sepsis.
Carraway
M S
MS
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710, USA.
Piantadosi
C A
CA
Jenkinson
C P
CP
Huang
Y C
YC
D000818:Animals; D001119:Arginase / Q000378:metabolism*; D015153:Blotting, Western; D004195:Disease Models, Animal; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000378:metabolism*; D008168:Lung / Q000201:enzymology* / Q000473:pathology; D008297:Male; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D009929:Organ Size; D009195:Peroxidase / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012128:Respiratory Distress Syndrome / Q000201:enzymology* / Q000473:pathology; D013047:Specific Pathogen-Free Organisms; D018746:Systemic Inflammatory Response Syndrome / Q000201:enzymology* / Q000473:pathology; D017392:Thiobarbituric Acid Reactive Substances
D007527:Isoenzymes; D017392:Thiobarbituric Acid Reactive Substances; D009195:Peroxidase; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D001119:Arginase