Reperfused Porcine Hearts Research Articles

Time delay of cell death by Na+/H+-exchange inhibition in regionally ischemic, reperfused porcine hearts.

Studies in different preparations have suggested that Na+/H+ exchange is one mechanism causally involved in cell death in myocardial ischemia and reperfusion. The time delay of cell death by pretreatment with the Na+/H+-exchange inhibitor HOE642, cariporide (4-isopropyl-3-methylsulphonylbenzoyl-guanidine methanesulphonate), was investigated in regionally ischemic, reperfused porcine hearts. HOE642 (1 mg/kg) was injected intravenously in 14 thoracotomized pigs 10 min before occlusion of the left anterior descending coronary artery (45 min of ischemia, six pigs; 70 min of ischemia, six pigs; 90 min of ischemia, two pigs). Ischemia was followed by 24 h of reperfusion. Six animals (45 min of ischemia) served as controls. Infarct size was determined as a ratio of infarcted (tetrazolium stain, histology) to ischemic myocardium (dye technique), and regional myocardial function was assessed by sonomicrometry. HOE642 did not affect global hemodynamic parameters. In the pretreated group with 45 min of ischemia, HOE642 significantly decreased histochemical infarct size from 51.2 +/- 12.6% (control group) to 13.2 +/- 12% (p < 0.005) and histologic infarct size from 44.5 +/- 9% to 17.1 +/- 7% (p < 0.005). Recovery of regional systolic shortening after 24 h of reperfusion was improved from 2 +/- 6% (control group) to 12 +/- 7% (p = 0.02). In addition, myocardial contracture and increase in heart rate during early reperfusion were attenuated. When ischemia was prolonged to 70 min after pretreatment with HOE642, infarct size, recovery of systolic shortening, myocardial contracture, and increase in heart rate did not differ from those of the control group. Pretreatment with HOE642 increased the tolerance to ischemia/reperfusion by approximately 20-25 min. Inhibition of Na+/H+ exchange appears to be very promising in the clinical treatment of acute myocardial ischemia and reperfusion.

Lethal Reperfusion Injury in Regionally Ischemic, Reperfused Porcine Hearts?

Lethal Reperfusion Injury in Regionally Ischemic, Reperfused Porcine Hearts?

1005-81 Myocardial Protection by Na+/H+ Exchange Inhibition in Ischemic, Reperfused Porcine Hearts

The protective effect of the Na + /H + exchange inhibitor HOE 694 was tested in porcine hearts subjected to 45 min of regional ischemia and 24 h of reperfusion. The compound (3 mg/kg) was intravenously injected in 6 pigs each either 10 min before ischemia (group A) or 10 min before reperfusion (group B). Six animals served as controls. Apart from the main end-points, infarct size and regional systolic shortening, the effect of HOE 694 on global hemodynamic parameters which included coronary blood flow and coronary venous oxygen saturation was evaluated. Although the Na + IH + exchange inhibitor did not affect global hemodynamics, preischemic treatment with HOE 694 decreased infarct size from 65 ± 18% (control group) to 12 ± 9% (p &lt; 0.01) and improved systolic shortening from 8 ± 6% (control group) to 28 ± 9% P &lt; 0.02). In addition, increase in heart rate and myocardial contracture during early reperfusion were significantly attenuated in group A. Treatment of group B did not exhibit protective effects. Na + /H + exchange inhibition is a very protective means in myocardial ischemia and reperfusion when administered before ischemia. In this model, it was ineffective when given before reperfusion.

Effects of R56865, an Na(+)- and Ca(2+)-overload inhibitor, on myocardial injury in ischemic, reperfused porcine hearts.

The cardioprotective effect of R56865, an Na(+)- and Ca(2+)-overload inhibitor, was studied in 20 regionally ischemic reperfused (I/R) porcine hearts. Ten pigs were treated with a single intravenous (i.v.) injection of 0.4 mg/kg 15 min before occlusion of the distal left anterior descending coronary artery (LAD) for 45 min. Ten other animals served as controls. Infarct size (IS) was determined as percentage of infarcted (tetrazolium method) to ischemic (dye technique) myocardium after 24-h reperfusion. Regional systolic shortening (SS) was determined by sonomicrometry. Fifteen minutes after i.v. administration of R56865, a significant decrease in heart rate (HR) (from 83 +/- 15 to 74 +/- 16 beats/min), dP/dtmax (from 2,033 +/- 604 to 1,822 +/- 524 mm Hg/s), LAD blood flow (BF, from 17 +/- 7 to 14 +/- 7 ml/min), and calculated global myocardial O2 consumption (MVO2) (from 5.9 +/- 0.9 to 5.4 +/- 0.9 ml O2/min x 100 g) was observed. Although this investigational drug attenuated the increase in HR during early reperfusion, the incidence of ventricular fibrillation (VF) was not affected during either ischemia or reperfusion. R56865 reduced IS by 24% from 67.1 +/- 16% (control group) to 50.8 +/- 13%. In addition, this treatment improved systolic shortening after 24-h reperfusion from 6 +/- 8% (control group) to 15 +/- 9%. Our results support the concept that inhibition of intracellular Na(+)- and Ca(2+)-overload is a promising new principle in treatment of myocardial I/R.

Influence of coronary venous retroinfusion and vasodilatation on regional myocardial blood flow measurement with microspheres. An analysis of 'microsphere loss' from ischaemic and reperfused porcine hearts.

The influence of coronary venous retroinfusion and a vasoselective calcium antagonist felodipine on the microsphere loss in a porcine model of myocardial ischaemia and reperfusion was studied. Sixteen open-chest pigs underwent 45 min of myocardial ischaemia induced by occlusion of the left anterior descending coronary artery followed by 4 h of reperfusion. Either felodipine (felo-retro group, 7 nmol kg-1, n = 6) or the corresponding amount of vehicle (vehicle-retro group, n = 5) was infused retrogradely into the coronary veins over 30 min, starting 5 min before reperfusion. In a third group, the same amount of felodipine was administered intravenously (felo-iv group, n = 5). Myocardial regional blood flow was measured with radiolabelled microspheres (phi = 15 microns) injected before ischaemia to investigate a possible loss during ischaemia. In the felo-retro group, the apparent blood flow in the ischaemic areas, expressed as a percentage of the corresponding values in the non-ischaemic areas (%-flow), were 73 +/- 15, 73 +/- 11 and 75 +/- 19 in the subendocardial, midmyocardial and subepicardial layers, respectively. The corresponding percentage flows were 64 +/- 11, 70 +/- 11 and 62 +/- 9 in the vehicle-retro group and 75 +/- 18, 77 +/- 15 and 76 +/- 11 in the felo-iv group. The differences between the groups were not statistically significant. It is concluded that in this open-chest preparation microsphere loss observed in the ischaemic and reperfused myocardium is not increased by coronary venous retroinfusion or by a concomitantly administered vasodilative agent like felodipine.

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 μg/min) and continued until 45 minutes of reperfusion (mean 18 μg/min). Immediately before and during 45 minutes of reperfusion, left ventricular peak pressure, dpdtmax, 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 β-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.

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

Ischemic, 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 mumol/mg frozen weight, treated group 1.7 +/- 0.4 mumol/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.

Stunning in partially infarcted, reperfused porcine hearts: Hermann H. Klein Dept. of Cardiology, University Hospital Lahnberge, Marburg, Germany

Stunning in partially infarcted, reperfused porcine hearts: Hermann H. Klein Dept. of Cardiology, University Hospital Lahnberge, Marburg, Germany

The effects of Trolox, a water-soluble vitamin E analogue, in regionally ischemic, reperfused porcine hearts

Myocardial protection by the water-soluble vitamin E analogue, Trolox, was investigated in 18 regionally ischemic, reperfused porcine hearts. The left anterior descending coronary artery was distally ligated for 45 min and was reperfused for three days. Five grams of Trolox ( n = 9) were infused intravenously before coronary occlusion. Treatment was continued with an intravenous dose of 5 grams Trolox/24 hours 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. Generation of free radicals by stimulated neutrophils was evaluated by luminol-enhanced chemiluminescence. Plasma concentrations of Trolox were measured by high-performance liquid chromatography. Aside from heart rate before ischemia, global hemodynamic values including calculated left ventricular oxygen consumption did not differ significantly between the two groups. Plasma concentrations of Trolox measured 1.8 ± 0.3 mmol/l (before ischemia), 0.96 ± 0.13 mmol/l (before reperfusion), 0.77 ± 0.1 mmol/l (40 min of reperfusion), and 0.08 mmol/l (end of the experiment). Generation of free radicals by stimulated neutrophils was reduced by about 30% in the treatment group before ischemia and immediately before reperfusion, but was not reduced at the end of the experiment. Risk regions (control group 19.4 ± 6 g, treatment group 19.3 ± 7 g) and infarct sizes (control group 69.3 ± 8%, treatment group 69.3 ± 12%) were almost identical. Regional systolic shortening of a control segment and of the risk region were similar in both groups before ischemia, before reperfusion, and after 45 min of reperfusion. After 3 days of reperfusion, regional systolic shortening of the reperfused myocardium of the treated group had recovered to a significantly greater extent ( P = 0.027). This parameter amounted to 9 ± 6% in the treated group and to 3 ± 3% in the control group. Improved functional recovery was not accompanied by higher tissue concentrations of adenosine triphosphate. It is concluded that the chosen treatment with Trolox does not reduce infarct size but accelerates functional recovery. This finding suggests that the mechanisms resulting in myocardial necrosis during ischemia/reperfusion and in post-ischemic myocardial dysfunction may differ.

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 ilchemia 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.

Combined treatment with vitamins E and C in experimental myocardial infarction in pigs

The effect of two different combined treatments with vitamin E acetate and vitamin C on infarct size and recovery of regional myocardial function was investigated in ischemic, reperfused porcine hearts. The left anterior descending coronary artery was distally ligated in 30 thoracotomized pigs for 45 minutes followed by 3 days of reperfusion. Infarct size was determined as the ratio of infarcted (tetrazolium stain) to ischemic (dye technique) myocardium. Regional myocardial function was assessed by sonomicrometry. Ten pigs received vitamin E acetate (12 gm intravenously three times for 1 week) before ischemia and vitamin C (4.4 gm intravenously) before reperfusion (therapy A). Another 10 pigs were treated with vitamin E acetate (12 gm intraarterially) and vitamin C (4.4 gm intravenously) during ischemia (therapy B). An additional 10 pigs served as a control group. Global hemodynamics did not differ significantly among the groups before and during ischemia. Mean plasma concentrations of vitamin E amounted to 107 μg/ml in group A, 16 μg/ml in group B, and 0.9 μg/ml in the control group at the onset of reperfusion. Therapy A reduced the size of the infarct from 73 ± 12% to 47 ± 16% of the region at risk ( p < 0.005) and improved regional systolic shortening from 0 ± 7% to 11 ± 6% at 3 days after reperfusion ( p < 0.01). Therapy B decreased the size of the infarct to 64 ± 9% of the region at risk ( p = 0.05). Regional systolic shortening was not significantly improved by this treatment protocol ( p = 0.18). We conclude that therapy A protects the ischemic, reperfused myocardium probably by scavenging free radicals formed during ischemia and/or during reperfusion. Results of this study do not indicate whether these radicals were generated by jeopardized myocytes or by activated inflammatory cells. Therapy B exhibited only borderline protection. These findings show that cell death of ischemic, reperfused myocytes can be reduced by appropriate treatment with pharmacologic doses of vitamin E acetate and vitamin C.

Treatment of reperfusion injury with intracoronary calcium channel antagonists and reduced coronary free calcium concentration in regionally ischemic, reperfused porcine hearts

The effect of intracoronary diltiazem, EGTA (ethylenebis-(β-aminomethylether)-N,N′-tetraacetic acid), nifedipine, verapamil and isotonic saline solution as placebo on reperfusion injury was investigated in regionally ischemic, reperfused porcine hearts. The left anterior descending coronary artery was distally occluded for 45 min and was reperfused for 3 days. Intracoronary infusion was started immediately before reperfusion and continued during 45 min of reperfusion. Infarct size was determined as the ratio of infarcted (tetrazolium stain) to ischemic myocardium (dye technique). Regional systolic shortening was assessed by sonomicrometry.Apart from left ventricular end-diastolic pressure before ischemia and during 45 min of reperfusion, global hemodynamic values in the five treatment groups did not differ; in particular, calculated left ventricular oxygen consumption before and during ischemia was equally low. Intracoronary EGTA decreased coronary venous free calcium concentration to about 70% of baseline value. Infarct size was reduced from 76 ± 10% (control group, n = 8) to 60 ± 10% (p < 0.01) by intracoronary diltiazem (n = 8) and to 55 ± 15% (p < 0.01) by intracoronary EGTA (n = 8). Insignificant reductions in infarct size were found after treatment with intracoronary verapamil (63 ± 18%, n = 8) and intracoronary nifedipine (68 ± 9%, n = 7). Regional systolic shortening of the risk region, which did not differ among the groups before occlusion and during ischemia, recovered to the greatest extent in the EGTA-treated pigs (p < 0.01 compared with values in the control group). Treatment with intracoronary calcium antagonists resulted in only marginal improvement of systolic shortening. Reperfusion-induced rhythm disturbances were not beneficially affected by either treatment.It is concluded that reperfusion injury exists in regionally ischemic, reperfused porcine hearts, which can be attenuated by intracoronary EGTA and intracoronary diltiazem administered during the first 45 min of reperfusion. The favorable action of EGTA is ascribed to a reduced ionized extracellular calcium concentration, whereas the beneficial mechanism of diltiazem is unknown.

The effect of intracoronary diltiazem on regional myocardial function and development of infarcts in porcine hearts.

The effect of intracoronary (i.c.) pretreatment with diltiazem on regional myocardial function and the development of infarcts was investigated in regionally ischemic, reperfused porcine hearts. The left anterior descending coronary artery (LAD) was distally ligated in 16 pigs for 20-90 min followed by 24 h of reperfusion. Eight pigs were treated with increasing doses of i.c. diltiazem (0.375 mg/min, 0.75 mg/min, 1 mg/min) prior to ischemia. Eight pigs served as controls. Regional myocardial function was assessed by implanted ultrasonic crystals. Infarct size was determined as ratio of infarcted (tetrazolium stain) to ischemic myocardium (dye technique). I.c. diltiazem mainly depressed early systolic shortening (isovolumetric contraction) and lengthening during the first half of diastole. Pretreatment with this calcium antagonist significantly delayed the development of infarcts. In control experiments, a mean infarct size of 74% was found after 45-min ischemia. At that time no infarction was observed in the treated group, where infarcts started to evolve after 60-min ischemia. It is concluded that the favorable action of i.c. diltiazem can mainly be ascribed to a reduced myocardial oxygen consumption at the onset of ischemia.

Cell death in ischemic, reperfused porcine hearts: a histochemical and functional study.

The temporal development of infarcts was histochemically and functionally determined in porcine hearts. In one series of experiments (22 pigs), the distal third of the left anterior descending coronary artery (LAD) was transiently occluded for periods between 20 and 90 min and was reperfused for another 24 h. At the end of the experiments, the infarcted myocardium of four tissue slices was determined with a tetrazolium stain and related to the risk region which was delineated by a fluorescent dye. Infarcts started to develop in the ischemic septum and the subendocardial layer of the free anterior wall between 20 and 35 min of ischemia. Thereafter, infarctions progressed rapidly from the inner towards the outer layer at risk. The jeopardized anterior left ventricular wall became almost completely infarcted within 60 min of ischemia. In a second series of experiments (10 pigs) recovery of systolic shortening was studied with implanted ultrasonic crystals over 3 weeks of reperfusion. At the end of the experiments, systolic shortening was about 75% of baseline level when ischemia had lasted between 20 and 35 min. Almost no recovery was observed when the occlusion time lasted 45 to 60 min. This study suggests that the assessment of myocardial infarction with a tetrazolium stain after 24 h of reperfusion corresponds very well with functional recovery after 3 weeks of reperfusion. Furthermore, determination of regional myocardial function of the ischemic, reperfused segment in the chronic stage may be considered an additional tool to evaluate therapeutic effects on infarct size in this model.

Verapamil does not reduce reperfusion injury in ischemic, reperfused porcine hearts

Verapamil does not reduce reperfusion injury in ischemic, reperfused porcine hearts

Intracoronary superoxide dismutase for the treatment of “reperfusion injury” A blind randomized placebo-controlled trial in ischemic, reperfused porcine hearts

The effect of recombinant human superoxide dismutase (rh-SOD) on infarct size was investigated in porcine hearts. The left anterior descending coronary artery was occluded in each of 24 anesthetized pigs for 45 min and reperfused for 24 h. The animals were randomly assigned to either rh-SOD (n = 12) or placebo treatment (n = 12). 2 min before reperfusion, an intracoronary (i.c.) infusion of rh-SOD (total dose: 2000 U/kg) or placebo was started which lasted for up to 45 min reperfusion. At the end of the experiment, the infarcted myocardium was assessed using a tetrazolium stain (NBT) and related to the risk region which was determined with a fluorescent dye. Two pigs of the SOD group and one of the control group died before the end of the experiments. Except for a lower calculated myocardial oxygen consumption and a lower dp/dtmax in the SOD group during ischemia, hemodynamic parameters of the two groups did not differ significantly. rh-SOD i.c. treatment during reperfusion did not reduce infarct size significantly. Infarct size amounted to 74 +/- 13% in the control group and to 66 +/- 19% in the treated group. The incidence of reperfusion arrhythmias was not affected by rh-SOD treatment. It is concluded that i.c. rh-SOD treatment at the beginning of reperfusion neither significantly reduces infarct size nor diminishes the incidence of reperfusion arrhythmias in this preparation.

Ischemic cell death in reperfused porcine hearts. A histochemical and fonctional study

Ischemic cell death in reperfused porcine hearts. A histochemical and fonctional study

Treatment with BW 755 C in ischemic, reperfused porcine hearts

Treatment with BW 755 C in ischemic, reperfused porcine hearts

Intracoronary hyperosmotic mannitol during reperfusion does not affect infarct size in ischemic, reperfused porcine hearts.

We investigated the effect of reperfusion with hyperosmotic mannitol on the infarct size in porcine hearts. The distal half of the left anterior descending coronary artery was occluded in each of 21 anesthetized pigs for 75 min and was reperfused for 2 h. During reperfusion mannitol (1075 mosmol/kg) was intracoronarily infused at a dose of 0.5 ml/min in 6 pigs ("low" mannitol group), at a dose of 1.5 ml/min in another 6 pigs ("high" mannitol group), and at a dose of 5 ml/min in 3 pigs for the first 8 min of reperfusion ("very high" mannitol group). 6 pigs served as controls. Although mannitol infusion increased plasma osmolality in the ischemic, reperfused myocardium in all experiments, the infarct size expressed as the ratio of the infarcted tissue over the area at risk of necrosis was not significantly influenced. Infarct size amounted to 72 +/- 25% in the control group, to 75 +/- 14% in the "low" mannitol group, to 78 +/- 18% in the "high" mannitol group, and to 93 +/- 8% in the "very high" mannitol group. These results clearly indicate that reperfusion with hyperosmotic mannitol after 75 min of ischemia does not exert any beneficial effect on the infarct size.

The effect of two different diltiazem treatments on infarct size in ischemic, reperfused porcine hearts.

The effect of diltiazem on the development of infarcts was investigated in porcine hearts. The left anterior descending coronary artery was occluded in each of 32 anesthetized pigs for 75 min and was reperfused for 4 hr. Diltiazem (15 micrograms/kg X min) was infused in 11 pigs for 30 min before occlusion (therapy A) and in another eight pigs before reperfusion (therapy B). Eleven pigs served as controls. Tissue concentrations of adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (NAD) were determined in transmural needle biopsy samples taken from the ischemic apex after 70 min of ischemia. The infarct size, expressed as the ratio of the infarcted tissue over the area at risk of necrosis multiplied by 100, amounted to 79 +/- 20% in the control group. Although there was no significant difference between hemodynamics in the control and the treated groups, pretreatment with diltiazem significantly reduced infarct size (53 +/- 26%; p = .025). Reduction of infarct size by therapy B did not reach the required level of significance (66 +/- 33%). The ischemic loss of ATP and NAD was significantly lower in the pretreated group, which further indicates that the beneficial effect of diltiazem was exerted primarily during ischemia and not during reperfusion.