Occurrence of oxidatively modified proteins: an early event in experimental acute pancreatitis.
Free radical-mediated injury is believed to play a key role in the pathogenesis of acute pancreatitis (AP). Therefore, oxidative damage of proteins may be an important event in the development of AP. The present study was performed to investigate oxidative protein modification, quantified as 2,4-dinitrophenylhydrazine-reactive protein-carbonyls, during the time course of taurocholate-induced pancreatitis of the rat and to analyze oxidatively modified proteins by Western blotting. Protein modification in pancreatic homogenates was found as early as 30 min after induction of severe AP with 3% taurocholate preceding the elevation of serum amylase activity and the increase of malondialdehyde in the tissue. A correlation of protein-carbonyl contents to a score of pancreatic macroscopic alterations (r = .69) and to the wet weight/dry weight ratio (r = .65) was found. Infusion of 5% taurocholate resulted in fulminant AP with high lethality during the 24 h of the experiment. However, rats surviving showed significantly lower level of protein-carbonyls than animals that died between 20-24 h after AP induction. The quantitative data were confirmed by the intensity of immunostained protein-carbonyls. The present data show a rather uniform increase in the staining pattern not revealing single, selectively damaged proteins. The aldehydic product of lipid peroxidation 4-hydroxynonenal (HNE) is known for its reactivity towards proteins. Interestingly, an antibody raised against protein-bound HNE did not indicate an increased protein modification by this aldehyde. In conclusion, experimental AP is characterized by an early oxidative protein modification, possibly contributing to functional impairment of the pancreas. This protein alteration may not be mediated by HNE.
Reinheckel
T
T
Centre of Surgery and Institute of Clinical Chemistry, Otto von Guericke University Magdeburg, Germany.
Nedelev
B
B
Prause
J
J
Augustin
W
W
Schulz
H U
HU
Lippert
H
H
Halangk
W
W
D000208:Acute Disease; D000447:Aldehydes / Q000737:chemistry; D000681:Amylases / Q000097:blood; D000818:Animals; D015153:Blotting, Western; D007700:Kinetics; D015227:Lipid Peroxidation; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D010084:Oxidation-Reduction; D010195:Pancreatitis / Q000139:chemically induced* / Q000378:metabolism; D010659:Phenylhydrazines / Q000737:chemistry; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D013656:Taurocholic Acid
D000447:Aldehydes; D010659:Phenylhydrazines; D011506:Proteins; C004787:2,4-dinitrophenylhydrazine; D008315:Malondialdehyde; D013656:Taurocholic Acid; D000681:Amylases; C027576:4-hydroxy-2-nonenal