Hepatic free radical production after cold storage: Kupffer cell-dependent and -independent mechanisms in rats.
OBJECTIVE
Free radicals are important mediators of reperfusion injury; however, the mechanism(s) of oxyradical production after liver reimplantation are not well understood. A model of cold storage and reperfusion using low-level chemiluminescence to directly measure oxyradical production during reperfusion was developed.


METHODS
Rat livers were harvested and stored at 4 degrees C in University of Wisconsin cold-storage solution or Euro-Collins solution for 0-48 hours and then flushed and reperfused with warm oxygenated (37 degrees C) Krebs-Henseleit buffer. Liver chemiluminescence was measured using a sensitive photomultiplier tube, and hepatocellular injury was assessed by measuring aspartate aminotransferase release into the perfusate.


RESULTS
Chemiluminescence reached a maximum within 5 minutes of reperfusion and then decreased to a baseline within 30 minutes. There was a marked increase in chemiluminescence after only a short period of storage in University of Wisconsin cold-storage solution. Chemiluminescence decreased with longer periods of storage but steadily increased again after 16 hours of storage. Chemiluminescence after 22 hours of storage, but not after 3 hours of storage, was decreased by pretreatment with the Kupffer cell inactivator gadolinium chloride.


CONCLUSIONS
The data suggest two mechanisms of oxyradical production during cold storage and reperfusion of the rat liver. The later phase seems to be Kupffer cell dependent.
Brass
C A
CA
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia.
Roberts
T G
TG
D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D001219:Aspartate Aminotransferases / Q000378:metabolism; D003080:Cold Temperature; D005609:Free Radicals; D005978:Glutathione; D006982:Hypertonic Solutions; D007328:Insulin; D007728:Kupffer Cells / Q000378:metabolism*; D016018:Least-Squares Analysis; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008163:Luminescent Measurements; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D010100:Oxygen / Q000378:metabolism*; D011887:Raffinose; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D015427:Reperfusion Injury / Q000378:metabolism
C037096:Euro-Collins' solution; D005609:Free Radicals; D006982:Hypertonic Solutions; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D001219:Aspartate Aminotransferases; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D010100:Oxygen