Oxidative stress and matrix metalloproteinase-9 activity in the liver after hypoxia and reoxygenation with 21% or 100% oxygen in newborn piglets.

We designed a randomized controlled study to identify and compare the liver tissue responses in systemic hypoxia and resuscitation with 21% and 100% oxygen using an animal model of neonatal hypoxia and reoxygenation. Twenty-seven piglets (1-3 days old, weight 1.5-2.0 kg) were acutely instrumented and mechanically ventilated. The animals underwent 2 h of normocapnic alveolar hypoxia (10-15% oxygen) then reoxygenation with 21% or 100% oxygen for 1 h, then 1 h with 21% oxygen. Controls were sham-operated without hypoxia-reoxygenation. After 2 h of reoxygenation liver tissue samples were immediately processed for histological and biochemical analyses of markers of oxidative stress and tissue injury. Two hours of hypoxia caused a significant reduction in mean arterial pressure with cardiogenic shock and metabolic acidemia, with similar recovery upon resuscitation with 21% and 100% oxygen. After 2 h of reoxygenation, the hepatic GSSG:total glutathione ratio and matrix metalloproteninase-9 activity, which correlated with the portal venous oxygenation at 15 min of reoxygenation, were greater in the 100% group and hepatic lactate level was higher in the 21% group than the controls (all P<0.05). Both hypoxic-reoxygenated groups had similarly elevated hepatic Bcl-2 levels. Apart from more non-distinct mitochondria identified in the 100% group, hepatic tissue adenylate energy charge and plasma transaminases levels did not differ among groups. We concluded that in this acute model of neonatal hypoxia and reoxygenation, resuscitation using 21% oxygen avoids the excess oxidative stress and elevated matrix metalloproteninase-9 activity in the liver when 100% oxygen was used. The study supports the conservative use of oxygen in optimizing post-hypoxic hepatic recovery.

Resuscitation with 21% or 100% oxygen is equally effective in restoring perfusion and oxygen metabolism in the liver of hypoxic newborn piglets.

The differential effects of the use of high or low oxygen levels during resuscitation on the neonatal liver are unknown. We compared the hepatic hemodynamics and oxygen metabolism in hypoxic newborn piglets resuscitated with 21% or 100% oxygen. Twenty-seven piglets (age, 1-3 days; weight, 1.5-2.0 kg) were acutely instrumented to measure cardiac output, hepatic artery, and portal venous blood flows (hepatic artery flow index [HAFI] and portal venous flow index [PVFI], respectively). The animals underwent 2 h of hypoxia (fraction of inspired oxygen, 0.10-0.15), then reoxygenation with 21% (n = 9) or 100% (n = 9) oxygen for 1 h, then 1 h with 21% oxygen. The controls (n = 9) were sham-operated without hypoxia-reoxygenation. Oxygen transport and plasma lactate concentrations were studied. Hypoxic animals had hypotension and decreased cardiac index with metabolic acidosis (mean pH, 7.00-7.02; P < 0.05 vs. controls). The PVFI and the total hepatic blood flow (THFI = PVFI + HAFI), despite the absence of significant change in HAFI, decreased to 16 +/- 2 mL/min/kg and 19 +/- 3 mL/min/kg, respectively (versus 24 +/- 2 mL/min/kg and 28 +/- 2 mL/min/kg of controls; P < 0.05). Fifteen minutes after reoxygenation, the cardiac index improved, PVFI recovered, HAFI was maintained, and THFI was not different between the groups. The hepatic oxygen consumption decreased (59%; P < 0.05) and the extraction increased (89%; P < 0.001) during hypoxia. Similarly, on reoxygenation, the hepatic oxygen consumption improved; however, extraction decreased versus controls on 100% but not on 21% oxygen (P < 0.05). The plasma lactate concentrations increased in both groups with hypoxia and were not different during reoxygenation between the group administered with 21% oxygen and the group administered with 100% oxygen. The hypoxic neonatal liver has reduced hepatic blood flow but has relatively preserved HAFI, and oxygen consumption recovered similarly on reoxygenation with 21% and 100% oxygen. The increased oxygen extraction during hypoxia normalized in 21% but reduced in 100% reoxygenation, with no differences in plasma lactate concentrations.

Platelet dysfunction in asphyxiated newborn piglets resuscitated with 21% and 100% oxygen.

Hemostatic disturbances are common in asphyxiated newborns after resuscitation. We compared platelet function in hypoxic newborn piglets reoxygenated with 21% or 100% oxygen. Piglets (1-3 d, 1.5-2.1 kg) were anesthetized and acutely instrumented for hemodynamic monitoring. After stabilization, normocapnic hypoxia was induced with an inspired oxygen concentration of 10-15% for 2 h. Piglets were then resuscitated for 1 h with 21% or 100% oxygen, followed by 3 h with 21% oxygen. Platelet counts and collagen (2, 5, and 10 microg/mL)-stimulated whole blood aggregation were studied before hypoxia and at 4 h of post-hypoxia/reoxygenation. Platelet function was studied using transmission electron microscopy and by measuring plasma thromboxane B2 (TxB2) and matrix metalloproteinase (MMP)-2 and -9 levels. Control piglets were sham-operated without hypoxia/reoxygenation. The hypoxemic (PaO2 33 mm Hg) piglets developed hypotension with metabolic acidosis (pH 7.02-7.05). Upon reoxygenation, piglets recovered and blood gases gradually normalized. At 4 h reoxygenation, platelet aggregation ex vivo was impaired as evidenced by a rightward-downward shifting of the concentration-response curves. Electron microscopy showed features of platelet activation. Plasma MMP-9 but not MMP-2 activity significantly increased. Resuscitation with 100% but not 21% oxygen increased plasma TxB2 levels. Platelet counts decreased after hypoxia/reoxygenation but were not different between groups during the experiment. Resuscitation of hypoxic newborn piglets caused platelet activation with significant deterioration of platelet aggregation ex vivo and increased plasma MMP-9 levels. High oxygen concentrations may aggravate the activation of prostaglandin-thromboxane mechanistic pathway.