Rapid increase in serum lipid peroxide 4-hydroxynonenal (HNE) through monocyte NADPH oxidase in early endo-toxemia.
We have developed a time-resolved fluoroimmunoassay (TR-FIA) for a lipid peroxide 4-hydroxynonenal (HNE), which is 100-fold more sensitive than conventional enzyme-linked immunosorbent assay (ELISA) and is an easier technique to use for a large number of samples without pre-treatment. By this assay, we found that a low dose of bacterial lipo-polysaccharide (LPS), injected intra-peritoneally (0.5 mg/kg), increased serum HNE level by 28-folds, with a peak at 20 min. LPS also increased HNE in vitro to a much higher level in the monocyte-enriched plasma than in the leukocyte-enriched plasma, with a peak at 10 min. The HNE production after LPS treatment was inhibited by apocynin, a specific NADPH oxidase inhibitor in vivo and in vitro, and to a lesser extent by dimethylsulfoxide a solvent for apocynin and a hydroxyl radical scavenger in vitro. These data suggest that monocyte NADPH oxidase is involved in the lipid peroxidation (HNE formation) in the LPS-challenged rat. This is the first clear demonstration of the link between an inflammatory stimulus and lipid peroxidation in the blood.
Kimura
Hiroko
H
Department of Forensic Medicine, Juntendo University School of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo, 113-8421, Japan.
Liu
Shuang
S
Yamada
Satoshi
S
Uchida
Koji
K
Matsumoto
Kazuko
K
Mukaida
Masahiro
M
Yoshida
Ken-Ichi
K
D000447:Aldehydes / Q000097:blood*; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000097:blood; D019446:Endotoxemia / Q000097:blood* / Q000201:enzymology; D015200:Fluoroimmunoassay / Q000379:methods; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000201:enzymology*; D008054:Lipid Peroxides / Q000097:blood*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008315:Malondialdehyde / Q000097:blood; D051379:Mice; D019255:NADPH Oxidases / Q000097:blood*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity
D000447:Aldehydes; D002352:Carrier Proteins; C030929:Crp protein, rat; D008054:Lipid Peroxides; D008070:Lipopolysaccharides; D008315:Malondialdehyde; D019255:NADPH Oxidases; C027576:4-hydroxy-2-nonenal