title: Xanthine oxidase released from reperfused hind limbs mediate kupffer cell activation, neutrophil sequestration, and hepatic oxidative stress in rats subjected to tourniquet shock.
abstract: We have shown previously that rats subjected to tourniquet shock develop an acute form of remote organ injury of the liver that is both Kupffer cell (KC) and polymorphonuclear (PMN) leukocyte dependent. Circulating plasma xanthine oxidase (XO) has been shown to be responsible for the development of endothelial dysfunction and for remote organ injury of the lung and intestine after ischemia-reperfusion protocols. We now hypothesize that XO is released from rat hind limbs upon reperfusion and that it is responsible for KC and PMN leukocyte activation in this shock model. Our results show that about 30% of rat gastrocnemius muscle xanthine dehydrogenase (XD) is converted to XO during the 5-h tourniquet period and that it is released into the femoral vein within 10 min of reperfusion. Total muscle xanthine oxidoreductase activity (XO + XD) decreases within 30 min of reperfusion and is paralleled by a corresponding increase in femoral vein lactic dehydrogenase. In addition, liver tissue XO increases significantly within 30 min of reperfusion without a corresponding conversion of endogenous XD. Conversion of hepatic XD becomes evident 60 min after reperfusion is initiated, as does XO, and alanine aminotransferase (ALT) release into the hepatic vein, presumably from damaged hepatocytes as a consequence of oxidative stress. Tissue myeloperoxidase activity also increases significantly after the 60-min reperfusion period. That XO mediates KC and PMN activation is supported by the following observations: a) the close relationships between plasma XO and the time courses of tumor necrosis factor-alpha TNFalpha release into the hepatic vein and colloidal carbon clearance by KCs; b) that colloidal carbon clearance, TNFalpha and ALT release, loss of tissue free thiols, lipid peroxidation (TBARS), and liver infiltration by PMN neutrophils can also be induced by the administration of exogenous XO to normal rats; and c) pretreatment of rats with allopurinol inhibits KC activation and liver leukocyte infiltration. These results suggest that XO, released from the ischemic limb on reperfusion, is taken up by the liver were it mediates KC and PMN neutrophil activation and thus contributes to the development of multiple system organ failure after hind limb reperfusion.
authors.affiliation: Departamento de Fisiopatología, Universidad de Concepción, Chile.
authors.forename: V L
authors.initials: VL
authors.lastname: Vega
authors.forename: L
authors.initials: L
authors.lastname: Mardones
authors.forename: M
authors.initials: M
authors.lastname: Maldonado
authors.forename: S
authors.initials: S
authors.lastname: Nicovani
authors.forename: V
authors.initials: V
authors.lastname: Manríquez
authors.forename: J
authors.initials: J
authors.lastname: Roa
authors.forename: P H
authors.initials: PH
authors.lastname: Ward
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D005260:Female; D006614:Hindlimb / Q000098:blood supply*; D007511:Ischemia / Q000503:physiopathology*; D007728:Kupffer Cells / Q000502:physiology*; D007770:L-Lactate Dehydrogenase / Q000097:blood; D008099:Liver / Q000502:physiology / Q000503:physiopathology*; D008262:Macrophage Activation; D018482:Muscle, Skeletal / Q000503:physiopathology; D009504:Neutrophils / Q000502:physiology*; D018384:Oxidative Stress / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015424:Reperfusion; D012769:Shock / Q000503:physiopathology*; D013997:Time Factors; D014111:Tourniquets; D014968:Xanthine Dehydrogenase / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism*
chemical_list: D007770:L-Lactate Dehydrogenase; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase; D000410:Alanine Transaminase