Endotoxin-induced aggravation of preservation-reperfusion injury of rat liver and its modulation.
OBJECTIVE
In clinical transplantation, exposure of donors to gut-derived endotoxin occurs frequently and may adversely affect liver transplantation therapy. The aim of this study was to investigate: 1) whether brief exposure of rats to endotoxin before liver procurement aggravates the early phase of reperfusion injury of hepatic explants; and if so 2) whether Kupffer cell activation is a contributing factor to liver injury; and 3) whether heparin and pentoxifylline could minimize this effect.


METHODS
Male Wistar rats were injected with 0.2-4.0 mg/kg of Escherichia coli lipopolysaccharide 2 h prior to liver harvest. After preservation in University of Wisconsin cold-storage solution, the livers were reperfused using a blood-free perfusion model. To inactivate Kupffer cells, some rats were pretreated with gadolinium chloride or liposome-encapsulated dichloromethylene-diphosphonate before lipopolysaccharide administration. The other rats received lipopolysaccharide with heparin or pentoxifylline.


RESULTS
In a dose-independent fashion, lipopolysaccharide impaired portal flow during graft reperfusion. In a dose-dependent way, lipopolysaccharide increased lactate dehydrogenase release into the perfusate and decreased bile flow and bromosulfophthalein excretion. Gadolinium chloride, liposomal dichloromethylene-diphosphonate, heparin, and pentoxifylline reduced lactate dehydrogenase release by 34%, 43%, 59%, and 64%, respectively, and improved functional parameters of the liver. A 52-fold increased neutrophil infiltration in the liver sinusoids after lipopolysaccharide exposure was not affected significantly by the drugs studied; however, heparin reduced markedly neutrophil activation.


CONCLUSIONS
The results of this investigation provide direct evidence that aggravation of preservation-reperfusion injury of rat liver by endotoxin is mediated by Kupffer cell-dependent mechanism(s) and it can be minimized by heparin and pentoxifylline.
Vajdová
K
K
Laboratory of Perfused Organs, Institute of Preventive and Clinical Medicine, Bratislava, Slovakia.
Smreková
R
R
Kukan
M
M
Jakubovský
J
J
van Rooijen
N
N
Horecký
J
J
Lutterová
M
M
Wsólová
L
L
D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D001646:Bile / Q000378:metabolism; D004002:Clodronic Acid / Q000494:pharmacology; D015925:Cryopreservation; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli; D005978:Glutathione / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007728:Kupffer Cells / Q000378:metabolism / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D009504:Neutrophils / Q000378:metabolism / Q000473:pathology; D009926:Organ Preservation; D019188:Organ Preservation Solutions; D010431:Pentoxifylline / Q000494:pharmacology; D010477:Perfusion; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism* / Q000517:prevention & control
D007328:Insulin; D008070:Lipopolysaccharides; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D004002:Clodronic Acid; D000493:Allopurinol; D006493:Heparin; D007770:L-Lactate Dehydrogenase; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D010431:Pentoxifylline