S. Pich

Failure of iloprost to protect the regionally ischemic, reperfused porcine heart

The effect of iloprost (Schering AG, Berlin), a stable prostacyclin analogue, was investigated in ischemic, reperfused porcine hearts. The left anterior descending coronary artery (LAD) was distally occluded in 18 pigs for 45 min followed by 3-d of reperfusion. Nine pigs were continuously treated with iloprost at a dose of 25 ng/kg per min. Treatment was started as intracoronary infusion into the proximal LAD 10 min before occlusion. The intercoronary infusion was replaced by an intravenous infusion after 45 min of reperfusion, which was continued until the end of the experiment. Infarct size was determined as the ratio of infarcted (tetrazolium stain) to ischemic myocardium (dye technique). Regional systolic shortening was assessed by sonomicrometry. Myocardial concentrations of adenosine triphosphate were evaluated at the end of the experiment. Generation of free radicals by stimulated polymorphonuclear neutrophils was determined by luminol-enhanced chemiluminescence. Histologic and immunohistologic techniques were applied to characterize the myocardial inflammatory response. Global hemodynamics did not differ between the two groups. Neither infarct size (control group 68 +/- 18%, treated group 74 +/- 14%), recovery of systolic shortening (control group 3 +/- 6%, treated group 6 +/- 6%), nor myocardial adenosine triphosphate concentrations were improved by iloprost treatment. Myocardial inflammatory response remained unaffected by this treatment. The capacity of coronary venous, stimulated polymorphonuclear neutrophils to generate free radicals was slightly suppressed in the treated group before ischemia, at the end of ischemia and during early reperfusion. In this preparation, iloprost did not exhibit any beneficial effect on infarct size, recovery of systolic shortening and myocardial adenosine triphosphate concentrations.

Acute treatment with vitamin E does not protect the regionally ischemic, reperfused porcine heart.

SummaryThirty pigs were randomly assigned to a blind treatment with vitamin E or placebo. Ten animals each received 0.5g d-alpha tocopherol intravenously before ischemia (group 1) or before reperfusion (group 2). Ten control pigs were treated with a lipid emulsion as placebo. The left anterior descending coronary artery was distally ligated for 45 min followed by 3 days of reperfusion. Infarct size was determined as ratio of infarcted (tetrazolium stain) to ischemic myocardium (dye technique). Regional systolic shortening was assessed by sonomicrometry. Myocardial and plasma concentrations of vitamin E were determined by high-performance liquid chromatography. Global hemodynamic parameters and estimated left ventricular oxygen consumption did not differ among the three groups. Intravenous treatment with vitamin E raised the plasma levels of this vitamin from 1 ± 0.3 mg/l (control group) to 21 ± 6 mg/l before ischemia, to 4 ± 2 mg/l before reperfusion and to 2 ± 0.6 mg/l at the end of the experiments in group 1. In group 2, vitamin E plasma levels increased from 1 ± 0.3 mg/l to 24 ± 13 mg/l before reperfusion and to 2 ± 0.6 mg/l after 3 days of reperfusion. At the end of the experiments, myocardial vitamin E concentrations amounted to 4.2 ± 0.7 ng/mg fresh weight (control group), 9.7 ± 2.1 ng/mg (group 1), and to 8.7 ± 1.4 ng/mg (group 2). The increase in vitamin E plasma concentration was not associated with a cardioprotective effect. Infarct sizes of the three groups (group 1: 68 ± 12%, group 2: 66 ± 15%, control group: 69 ± 8%) were almost identical. Furthermore, recovery of systolic shortening was not improved by the acute vitamin E treatment. Mean systolic shortening of the reperfused segment amounted to 4% in the two treatment groups and 3% in the control group after 3 days of reperfusion. These results suggest that an acute increase in vitamin E plasma concentration before ischemia or during the early phase of reperfusion does not protect the ischemic, reperfused porcine heart.

Postischemic cell death in reperfused porcine hearts is not attenuated by the spin trap agent PBN during early reperfusion

SummaryIschemic, reperfused porcine hearts were used to investigate whether the spin trap agent PBN (N-tert-butyl-alpha-phenylnitrone) attenuates postischemic cell death by scavenging of free radicals. The left anterior descending coronary artery (LAD) was ligated distally in 16 pigs for 45 min and then reperfused for 3 h. PBN (coronary concentration approximately 1 mM) was infused into the LAD of eight pigs during the first 45 min of reperfusion. Electron spin resonance spectroscopy (ESR) was performed to identify free radical adducts in the reperfused coronary venous blood. Regional systolic shortening (SS%) was determined by sonomicrometry. Infarct size was evaluated as the percentage of infarcted (tetrazolium stain) to ischemic (dye technique) myocardium. The transmural ultrastructural degree of myocardial injury as well as myocardial ATP levels were assessed at the end of the experiment.Intracoronary treatment with PBN during early reperfusion did not attenuate myocardial damage. Infarct sizes (control group 59±19%, treated group 55±14%), transmural ultrastructural alterations, myocardial ATP concentrations (control group 1.8±0.3 μmol/mg frozen weight, treated group 1.7±0.4 μmol/mg) and regional systolic shortening at the end of the experiments (control group −1±5%, treated group −2±6% did not differ significantly. Furthermore, under various experimental conditions of spin trapping, free radical adducts could not be identified in coronary venous blood during early reperfusion. The results suggest that the spin trap agent PBN (1 mM) does not affect postischemic cell death in porcine hearts.

Effect of epinephrine treatment during late ischemia and early reperfusion on regional myocardial function and infarct size in partially infarcted reperfused porcine hearts

This study investigated whether epinephrine treatment during late ischemia and early reperfusion improves systolic shortening after 45 minutes of reperfusion at the cost of increased infarct size. A model consisting of both stunned and dead myocytes was used. The left anterior descending coronary arteries of 10 control and 10 treated pigs were occluded distally for 40 minutes and then reperfused for 3 days. Regional systolic shortening was determined by sonomicrometry, and infarct size was assessed as the percentage of infarcted (tetrazolium stain) to ischemic (dye technique) myocardium. Intravenous administration of epinephrine was started 10 minutes before the onset of reperfusion (5 micrograms/min) and continued until 45 minutes of reperfusion (mean 18 micrograms/min). Immediately before and during 45 minutes of reperfusion, left ventricular peak pressure, dp/dtmax, and heart rate were significantly increased in the treated animals. After 45 minutes of reperfusion, epinephrine treatment improved systolic shortening of the reperfused myocardium (treated group 9% +/- 8%; control group -1% +/- 6%; p < 0.01). Transient beta-adrenergic stimulation of the reperfused myocardium did not increase infarct size (treated group 57.2% +/- 19%; control group 55.4% +/- 17%). In conclusion, epinephrine treatment during late ischemia and early reperfusion improved systolic shortening after 45 minutes of reperfusion without affecting infarct size.

Ultrastructural Evaluation of Postischemic Cell Death (Lethal Reperfusion Injury) in Porcine Hearts.

This study investigated whether reperfusion results in an increase of ultrastructurally determined myocardial injury in pig hearts. The left anterior descending coronary artery (LAD) was distally occluded in 12 pigs for 35–45 minutes and then reperfused for 3 hours. At the end of ischemia, as well as after 3 hours of reperfusion, one transmural biopsy was removed from the center of the risk region and subdivided into four specimens, representing the subendocardial (I), subendo-midmyocardial (II), subepi-midmyocardial (III), and subepicardial layers (IV). The degree of injury was assessed by electronmicroscopy and was scored as reversible (1), an almost equal mixture of reversible and irreversible (2), and totally irreversible (3) damage. In addition, infarct size was determined as the ratio of infarcted (tetrazolium stain) to ischemic (dye technique) myocardium. Infarct sizes ranged from 29.3% to 93% (mean 61.2%). The scores of injury of the four tissue layers before and after reperfusion did not differ significantly: layer I, 2.4 ± 0.8/2.3 ± 0.9; layer II, 2.2 ± 0.9/2.0 ± 0.9; layer III, 1.8 ± 0.9/2.0 ± 0.9; and layer IV, 1.6 ± 0.9/1.3 ± 0.6. The means of the four layers were almost identical at the end of ischemia (2.1 ± 0.8) and after 3 hours of reperfusion (2.0 ± 0.6). A linear regression analysis with 95% confidence limits of the score values before and after reperfusion indicated that maximally 25% of a mean final infarct size of about 50% may be due to lethal reperfusion injury. This study suggests that cell death in regional ischemia and reperfusion occurs predominantly during ischemia and not during reperfusion.