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Biomedical subjects

G J Gross

Publications and source records attributed to G J Gross.

At least 19 recordsLinked to original sources
PubMed1997

Increased tolerance of the chronically hypoxic immature heart to ischemia. Contribution of the KATP channel.

BACKGROUND: Hypoxia from birth in immature rabbits increases the tolerance of isolated hearts to ischemia compared with age-matched normoxic rabbits. We determined whether this increased tolerance to ischemia was due to an alteration in the ATP-sensitive potassium (KATP) channel and whether increased KATP channel activation was associated with increases in intracellular lactate. METHODS AND RESULTS: Isolated immature rabbit hearts (7 to 10 days old) were perfused with bicarbonate buffer at 39 degrees C in the Langendorff mode at a constant pressure. Saline-filled latex balloons were placed in the left and right ventricles for measurement of developed pressure. A KATP channel agonist (bimakalim) or a KATP channel antagonist (glibenclamide) was added 15 minutes before a global ischemic period of 18 minutes, followed by 35 minutes of reperfusion. Rabbits raised from birth in hypoxic conditions (FIO2 = 0.12) displayed significantly enhanced recovery of developed pressure. The right ventricle was more tolerant of ischemia than the left ventricle in normoxic and hypoxic hearts. Bimakalim (1 mumol/L) increased the recovery of left ventricular developed pressure in normoxic hearts to values not different from those of hypoxic controls (43 +/- 3% to 67 +/- 5%) and slightly increased developed pressure in hypoxic hearts (67 +/- 5% to 72 +/- 5%). Glibenclamide (3 mumol/L) abolished the cardioprotective effect of hypoxia (67 +/- 5% to 43 +/- 5%). Constant-flow studies indicated that the effects of bimakalim and glibenclamide were independent of their actions on coronary flow. Ventricular lactate and lactate dehydrogenase concentrations were elevated in hypoxic hearts compared with normoxic control hearts. CONCLUSIONS: Increased tolerance to ischemia exhibited by chronically hypoxic rabbit hearts is associated with increased activation of the KATP channel. This increased KATP activity may be the result of increased intracellular concentrations of lactate.

Analysis of Variance↗
PubMed1997

The ischemia-selective KATP channel antagonist, 5-hydroxydecanoate, blocks ischemic preconditioning in the rat heart.

Although the KATP channel has been demonstrated to be involved in ischemic preconditioning (IPC) in most species, controversy still exists as to the role of this channel as a mediator of PC in the rat heart. Previously, the authors' laboratories have shown that glibenclamide blocks IPC in the intact rat heart, in a time-dependent manner; however, since glibenclamide has been shown to have non-selective effects unrelated to KATP channel blockade, a structurally dissimilar and ischemia-selective KATP channel blocker, 5-hydroxydecanoate (5-HD), was used to further elucidate the role of KATP channels in mediating IPC in the rat heart. Anesthetized, open-chested Sprague-Dawley rats were subjected to one of four protocols. In Group I, control (C), rats were subjected to 30 min of left coronary artery occlusion and 90 min of reperfusion. In Group II, IPC was elicited by 1 x 5 min occlusion followed by 10 min of reperfusion, prior to the 30 min occlusion and 90 min reperfusion periods, 5-HD (5 mg/kg, i.v.) was given 15 min prior to the 30 min occlusion period in non-preconditioned animals, or given 15 min prior to IPC (5-HD+IPC) (Groups III and IV, respectively). Infarct size (IS), as a percentage of the area at risk (AAR), was determined by triphenyltetrazolium staining. Ischemic preconditioning produced a marked reduction in infarct size (47.5 +/- 3.8% to 7.9 +/- 1.9%, *P < 0.01), which was completely abolished by 5-HD (50.5 +/- 2.6%). These data further suggest that the opening of KATP channels is an important component of IPC in the intact rat heart, similar to that observed in other species.

Adenosine Triphosphate↗
PubMed1997

KATP channel activation in a rabbit model of chronic myocardial hypoxia.

Increased tolerance to ischemia exhibited in chronically hypoxia immature rabbit hearts is associated with increased activation of ATP-sensitive potassium (KATP) channels. We determined whether exposure to hypoxia from birth alters the electrophysiological characteristics of Purkinje fibers obtained from rabbits (n = 12/group) which were raised in a normoxic (F O2 = 0.21) or hypoxic (F1O2 = 0.12) environment from birth to 9 days of age and the involvement of the KATP channel. The endocardial surface was exposed and impaled with microelectrodes to record action potential characteristics from Purkinje fibers under control conditions and following exposure to glibenclamide (3 microM). Action potential durations (APD)90 in Purkinje fibers were significantly shorter in hypoxic hearts compared with normoxic controls (110 +/- 5 ms v 121 +/- 4 ms). Glibenclamide increased APD90 in hypoxic hearts (120 +/- 4 ms) to values similar to those observed in normoxic controls (121 +/- 4 ms). Glibenclamide had no effect on APD90 in normoxic hearts. Maximum diastolic potential was more negative in hypoxic hearts and this effect was attenuated by glibenclamide. We conclude that chronic myocardial hypoxia results in a shorter APD as compared with normoxic controls by enhanced activation of KATP channels.

Action Potentials↗
PubMed1997

Role of adenosine in isoflurane-induced cardioprotection.

BACKGROUND: This investigation tested the hypothesis that adenosine (A1) receptor blockade modulates the cardioprotective effects of isoflurane. METHODS: Hemodynamics and percentage segment shortening (%SS) in the left anterior descending coronary artery (LAD) perfusion territory were evaluated in barbiturate-anesthetized dogs (n = 31) at selected intervals after pretreatment with the selective A1 receptor antagonist (8-cyclopentyl-1,3,dipropyl-xanthine; DPCPX 0.8 mg/kg, intravenously) or drug vehicle in the presence or absence of 1 minimum alveolar concentration (MAC) isoflurane. Dogs were subjected to five 5-min occlusions and reperfusions of the LAD, followed by 180 min of final reperfusion. Isoflurane was administered for 30 min before and during LAD occlusions and reperfusions and was discontinued at the onset of final reperfusion. Two other groups of dogs (n = 17) were used to measure interstitial concentrations of purines in the LAD region using a microdialysis technique in the presence and absence of isoflurane. RESULTS: Dogs receiving drug vehicle or DPCPX exhibited no recovery of %SS after 180 min of reperfusion (-5 +/- 7 and 5 +/- 11% of baseline, respectively, +/- SEM). In contrast, dogs receiving isoflurane alone demonstrated complete recovery of %SS at 60 min after reperfusion. DPCPX pretreatment partially attenuated isoflurane-induced enhancement of recovery of %SS (34 +/- 11% of baseline 180 min after reperfusion; P < 0.05). Interstitial purine concentrations were increased during multiple occlusions and reperfusions of the LAD in dogs not receiving isoflurane, but they were unchanged by coronary artery occlusion and reperfusion in dogs receiving isoflurane. CONCLUSIONS: The results indicate that isoflurane-induced cardioprotection in stunned myocardium is partially mediated by adenosine type 1 receptor activation and is accompanied by decreases in endogenous adenosine release.

Anesthetics, Inhalation↗
PubMed1997

KATP channels and memory of ischemic preconditioning in dogs: synergism between adenosine and KATP channels.

Results from numerous studies have shown that there is an important link between adenosine A1 receptors and ATP-sensitive potassium (KATP) channels in mediating the cardioprotective effects of ischemic preconditioning (PC). The major aim of the present study was to determine whether occupation of A1 receptors and/or the opening of KATP channels is involved in the time delay between the PC stimulus and the prolonged ischemic insult or the "memory" of PC to reduce infarct size. Barbital sodium-anesthetized dogs were subjected to 1 h of left anterior descending coronary artery (LAD) occlusion followed by 4 h of reperfusion. Ischemic PC was elicited by 10 min of LAD occlusion followed by 1 h of reperfusion (1-h memory) before the 1-h occlusion period. Either adenosine (800 g/min), bimakalim (3 g/min), a combination of two lower doses of each agent (400 g/min of adenosine and 0.3 g/min of bimakalim), or an equivalent volume of saline was infused into the LAD for 10 min followed by a 1-h drug-free period before the 1-h ischemic insult. In another series, glibenclamide, 8-cyclopentyl-1,3-dipropylxanthine (a selective A1-receptor blocker), or PD-115199 (a nonselective adenosine-receptor antagonist) was administered 50 min after ischemic PC (10 min before the 1-h occlusion period). Infarct size (IS) was expressed as a percentage of the area at risk. PC with 1 h of reperfusion resulted in a marked reduction in IS (8.1 +/- 6.5 vs. 29.8 +/- 5.8% in control dogs). Administration of adenosine or bimakalim followed by a 1-h drug-free period had no effect on IS; however, the simultaneous administration of adenosine and bimakalim resulted in a marked decrease in IS (11.5 +/- 2.7%). One hour after ischemic PC, administration of glibenclamide blocked the protective effect of ischemic PC, whereas 8-cyclopentyl-1,3-dipropylxanthine or PD-115199 did not affect it. These results provide evidence that the opening of myocardial KATP channels may play an important role in the memory of ischemic PC in the canine heart and also suggest that adenosine and the KATP channel may have a synergistic interaction that is important for the memory phase of PC.

Adenosine↗
PubMed1997

Adenosine A1 receptor blockade does not abolish the cardioprotective effects of the adenosine triphosphate-sensitive potassium channel opener bimakalim.

There has been controversy regarding whether ATP-sensitive potassium channel activation protects hearts through adenosine A1 receptor activation or the converse. We addressed this issue by determining the effect of the adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) on the cardioprotective activity of the ATP-sensitive potassium channel opener bimakalim. In isolated rat hearts subjected to 25 min of global ischemia and 30 min of reperfusion, bimakalim significantly reduced lactate dehydrogenase release and improved postischemic recovery of contractile function. Bimakalim increased the time to the onset of ischemic contracture (EC25 = 1.2 microM), compared with vehicle, and 10 microM DPCPX had no effect on this protective action (EC25 = 1.1 microM). The 10 microM concentration of DPCPX was sufficient to abolish the bradycardic and cardioprotective effects of the adenosine A1 receptor agonist (R)-(-)-N6-(2-phenylisopropyl)adenosine. DPCPX alone had no effect on the severity of ischemia/reperfusion damage. Glyburide completely abolished the cardioprotective effects of bimakalim. Bimakalim (1 microg/kg, intracoronarily) given over four periods of 5 min, interspersed with 10-min drug-free periods, before a 60-min occlusion and 3-hr reperfusion significantly reduced infarction size in anesthetized dogs (25 +/- 5 and 8 +/- 2% of the left ventricular area at risk for vehicle- and bimakalim-treated groups, respectively). DPCPX had no effect on the infarction-sparing activity of bimakalim (9 +/- 3% of the left ventricular area at risk). The protective effect of bimakalim was not accompanied by marked hemodynamic changes or by changes in regional myocardial blood flow. The results of this study suggest that the cardioprotective effects of ATP-sensitive potassium channel openers are not dependent on adenosine A1 receptor activation in rat or dog models of ischemia.

Animals↗
PubMed1996

Antibody to platelet/endothelial cell adhesion molecule-1 reduces myocardial infarct size in a rat model of ischemia-reperfusion injury.

BACKGROUND: Antibodies to selected neutrophil or endothelial cell adhesion molecules decrease myocardial infarct size in vivo. Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is an immunoglobulin gene superfamily member expressed constitutively on neutrophils and endothelium. F(ab')2 fragments of antibody against PECAM-1 inhibit transendothelial migration of neutrophils in several in vivo models of acute inflammation. Therefore, we examined the effect of F(ab')2 fragments of anti-PECAM-1 antibody in a rat model of myocardial infarction. METHODS AND RESULTS: F(ab')2 fragments of the anti-PECAM-1 antibody SEW16 and control normal rabbit IgG (NRIgG) were administered at 5 mg/kg to male Wistar rats, and the rats were subjected to a 30-minute coronary artery occlusion followed by 2 hours of reperfusion. At the completion of each experiment, the area at risk, infarct size (IS), and myeloperoxidase (MPO) activity were determined. Compared with untreated (n = 8; IS, 57 +/- 5%) or NRIgG-treated (n = 10; IS, 62 +/- 3%) control rats, SEW16-treated rats (n = 15; IS, 28.5 +/- 4%) displayed a 54% decrease in myocardial infarct size (P < .001). Hemodynamic parameters, leukocyte counts, total left ventricular weight, and area-at-risk weights did not differ significantly between the treatment groups. However, measurement of MPO activity revealed that neutrophil accumulation was reduced 83% (NRIgG, 975 +/- 55 mU/g; SEW16, 167 +/- 62 mU/g). CONCLUSIONS: These results demonstrate that blocking PECAM-1 exerts a significant protective effect in a rat model of myocardial ischemia-reperfusion injury via blockade of neutrophil accumulation in the myocardium.

Analysis of Variance↗
PubMed1996

Simultaneous determination of adenosine, inosine, hypoxanthine, xanthine, and uric acid in microdialysis samples using microbore column high-performance liquid chromatography with a diode array detector.

In the myocardial interstitial space, adenosine and its metabolites are important markers of ischemia, regulators of blood flow, and may produce cardioprotection against ischemia. A fast and sensitive method to assess the concentrations of adenosine and its metabolites is necessary to determine their involvement in mediating these effects. A method for the simultaneous determination of adenosine, inosine, hypoxanthine, xanthine, and uric acid in the interstitial fluid of the canine myocardium was developed using microdialysis, microbore column high-performance liquid chromatography, and a photo diode array detector (DAD). The microdialysis samples were injected directly onto a microbore C18 reverse-phase column without any prior sample preparation. Use of a DAD in this method provided many advantages. First, a DAD allowed the simultaneous detection of UV absorbance at multiple wavelengths, allowing the detection of each compound at their maximal UV absorbance. Further, the full UV absorption spectrum was recorded for each detected peak, confirming peak purity and identity. Using a microbore HPLC column and detection of UV absorbance at the maximal absorbance for each compound improve the sensitivity for all compounds. The detection limit of these compounds is 50 fmol (signal-to-noise ratio, S/N = 3). This method is useful in analyzing the temporal effect of a prolonged period of myocardial ischemia and reperfusion upon interstitial adenosine, inosine, hypoxanthine, xanthine, and uric acid concentrations in an in vivo canine model.

Adenosine↗
PubMed1996

Infarct size-reducing effect of nicorandil is mediated by the KATP channel but not by its nitrate-like properties in dogs.

OBJECTIVES: We wished to determine whether the cardioprotective effect of nicorandil to reduce infarct size is blocked by glibenclamide, a selective KATP channel antagonist, or methylene blue, a nitric oxide (NO)/guanylate cyclase inhibitor, in dogs. The second aim was to determine if glyceryl trinitrate produces a cardioprotective effect in the same model and to test if this effect is blocked by methylene blue and not by glibenclamide. We also determined whether adenosine release from the ischemic-reperfused area is an accurate index of ischemic severity in the presence of these drugs. METHODS: Barbiturate-anesthetized dogs were subjected to 60 min of left anterior descending coronary artery (LAD) occlusion followed by 3 h of reperfusion. In the first three groups, either nicorandil (100 micrograms/kg bolus + 10 micrograms/kg/min), glyceryl trinitrate (10 micrograms/kg bolus + 1 microgram/kg/min) or an equivalent volume of saline was given intravenously 15 min before LAD occlusion and continued to the time of reperfusion. In the next three groups, glibenclamide (0.3 mg/kg) was administered 15 min before drug infusion. In the final three groups, methylene blue (80 microM) was given intracoronarily 5 min before nicorandil or glyceryl trinitrate and continued until 15 min following reperfusion. Coronary venous blood samples were collected at various times during ischemia and following reperfusion and the concentration of adenosine measured. RESULTS: Nicorandil produced a marked reduction in infarct size expressed as a percent of the area at risk (NC group, 12.2 +/- 3.2% vs. Control group, 25.7 +/- 4.1%, P < 0.05) and this effect was completely abolished by pretreatment with glibenclamide. However, intracoronary administration of methylene blue did not block the cardioprotective effect of nicorandil. On the other hand, glyceryl trinitrate also produced a significant reduction in infarct size (GTN group, 13.0 +/- 3.1%) and this effect was reversed by methylene blue but not by glibenclamide. Adenosine concentrations in coronary venous blood were significantly reduced after reperfusion in the groups with small infarctions as compared with the Control group. CONCLUSIONS: These results suggest that at equieffective cardioprotective doses the infarct size-reducing effect of nicorandil in dogs is mediated via opening of myocardial KATP channels and that the cardioprotective effect of glyceryl trinitrate is most likely to be mediated via activation of guanylate cyclase at a site yet to be determined.

Adenosine↗
PubMed1996

Predictive value of proximal femoral bone densitometry in determining local orthogonal material properties.

Models which are based on non-invasive bone measurements may in the future be able to successfully identify individual subjects at an increased risk for hip fracture; thus, we designed a study to determine the usefulness of dual-energy X-ray absorptiometry (DXA) and quantitative computed tomography (QCT) in predicting the local material properties of human proximal femoral cancellous bone. There has been some disagreement in the scientific literature regarding appropriate predictive models for local material properties of cancellous bone. We sought to confirm that density-mechanical property relationships were consistent from subject to subject, and that three-dimensional QCT measurements were stronger predictors of mechanical properties than two-dimensional DXA results. Linear and power fit relationships between these densitometric measures and material properties were also examined to determine which were more appropriate. Bone cubes from specific regions of highly oriented trabeculae were analyzed separately to determine if cube orientation had an effect on mechanical properties independent of bone density. Ten pairs of ex vivo femurs (five male, five female; age 30-93, mean age 62) were prepared such that specific anatomic planes were visible radiographically. Both QCT and DXA measurements were made on all 20 femurs. Cancellous bone cubes were obtained proceeding along two distinct directions from the proximal end of each femur pair. Unexpectedly, the density-modulus relationships among these ten donors were found to be significantly different at p <0.01 (83 percent of the tests were different at p <0.0001). Density-strength regressions were also significantly different at p <0.01, but this effect was not as consistent nor as statistically significant. In general, the QCT method did not produce predictions of local cancellous bone material properties superior to the DXA method. The linear and power fit models appeared to produce consistent results, with neither being obviously more advantageous. These density measurements explained at best 30-40 percent of the variance in modulus and 50-60 percent of the variance in ultimate stress. The orientation of cancellous cubes in the principal compressive trabeculae region was a significant contributor to mechanical properties (p= 0.0001) independent of bone density. This finding was not as dramatic in the femoral neck cancellous bone region.

Absorptiometry, Photon↗
PubMed1996

Glyburide, a KATP channel antagonist, attenuates the cardioprotective effects of isoflurane in stunned myocardium.

This investigation examined the role of myocardial adenosine triphosphate-regulated potassium (KATP) channels in isoflurane-induced enhancement of myocardial function after reversible tissue injury produced by a 15-min left anterior descending coronary artery occlusion (LAD) and reperfusion. Dogs (n = 14) were chronically instrumented for measurement of left ventricular (LV) and aortic blood pressure, cardiac output, coronary blood flow velocity, and subendocardial segment length. Regional myocardial contractility was evaluated with preload recruitable work area (PRWA). Isovolumic relaxation was assessed with a time constant (tau). Hemodynamic variables and LV function were measured in the conscious state, during 2% isoflurane anesthesia for 45 min before and during a 15-min LAD occlusion, and at several intervals after reperfusion in dogs pretreated with glyburide (0.3 mg/kg, intravenously) or drug vehicle. LAD occlusion caused regional dyskinesia and increases in tau. Vehicle-pretreated dogs demonstrated full recovery of segment shortening by 5 h postreperfusion and recovery of PRWA and tau by 30 min postreperfusion. In contrast, dogs pretreated with glyburide demonstrated sustained systolic and diastolic dysfunction. Segment shortening recovered to only 70% +/- 5%, PRWA remained depressed at 48% +/- 10% and tau was prolonged to 116% +/- 5% of control values at 5 h postreperfusion. The results indicate that isoflurane enhances recovery of myocardial contractile function by 5 h postreperfusion, in comparison to previous findings in conscious dogs. These effects are partially blocked by glyburide pretreatment, indicating that KATP channel activation by isoflurance may mediate these cardioprotective effects.

Anesthetics, Inhalation↗
PubMed1996

Mechanism of myocardial protection by isoflurane. Role of adenosine triphosphate-regulated potassium (KATP) channels.

BACKGROUND: The mechanism of the protective actions of volatile anesthetics in ischemic myocardium has not been clearly elucidated. The role of myocardial adenosine triphosphate-regulated potassium (KATP) channels in isoflurane-induced enhancement of recovery of regional contractile function after multiple brief occlusions and reperfusion of the left anterior descending coronary artery (LAD) was studied in dogs anesthetized with barbiturates. METHODS: Dogs (n = 32) were instrumented to measure left ventricular and aortic blood pressure, cardiac output, LAD coronary blood flow velocity, and subendocardial segment length. Regional myocardial perfusion was measured using radioactive microspheres. Hemodynamics and percentage segment shortening (%SS) in the LAD perfusion territory were evaluated after instrumentation was complete; after pretreatment with the KATP channel antagonist, glyburide (0.05 mg/kg-1) or drug vehicle (polyethylene glycol in ethyl alcohol; control experiments); and in the presence or absence of 1 MAC isoflurane administered for 30 min before and during five 5-min occlusions and reperfusion of the LAD in four experimental groups. Isoflurane was discontinued at the onset of the final reperfusion period. Measurements of hemodynamics, %SS, and myocardial perfusion were repeated at several intervals during 180 min after reperfusion of the LAD. RESULTS: Left anterior descending coronary artery occlusion caused regional dyskinesia during each 5-min occlusion in each dog. Control and glyburide-pretreated dogs demonstrated poor recovery of %SS by 180 min after reperfusion (2 +/- 10 and 7 +/- 6% of baseline, respectively). In contrast, dogs anesthetized with isoflurane exhibited complete recovery of function (%SS) by 180 min after reperfusion (82 +/- 8% of baseline). Enhanced recovery of regional contractile function by isoflurane was abolished by pretreatment with glyburide 180 min after reperfusion (16 +/- 10% of baseline). Improvement of functional recovery of stunned myocardium by isoflurane, and the blockade of this action by glyburide, was not associated with changes in hemodynamics or regional myocardial perfusion. CONCLUSIONS: The results indicate that isoflurane prevents decreased systolic shortening caused by multiple episodes of ischemia and reperfusion. These actions result in improved recovery of contractile function of postischemic, reperfused myocardium and are mediated by isoflurane-induced activation of KATP channels.

Adenosine Triphosphate↗
PubMed1996

One-day cold perfusion of bimakalim and butanedione monoxime restores ex situ cardiac function.

Bimakalim (Bim), an opener of ATP-sensitive K+ (KATP) channels, was given alone or with 2,3-butanedione monoxime (BDM), a reversible uncoupler of contractility, to protect myocardial function during 1 day of hypothermia. Left ventricular pressure (LVP), coronary flow (CF), percent O2 extraction (%O2E), and cardiac efficiency were measured in 96 isolated, perfused guinea pig hearts divided into seven groups: 1) cold control (no drugs); 2) BDM; 3) Bim; 4) BDM + Bim; 5) BDM + glibenclamide (Glib, a blocker of KATP channels); 6) BDM + Bim + Glib; and 7) time control (6 h warm perfusion only). Drugs were given before, during, and initially after 22 h of low CF at 3.8 degrees C. At 26 h (cold groups) or 4 h (warm group) LVP (mmHg; means +/- SE) was similar for time control (94 +/- 4) and BDM + Bim (92 +/- 4) groups, lower and equivalent in the BDM (65 +/- 7) and BDM + Bim + Glib (64 +/- 7) groups, but LVP was higher than in the Bim group (46 +/- 3), and lowest in the cold control (30 +/- 8) group. In addition, only in the BDM + Bim group were basal CF, %O2E, and cardiac efficiency returned to values obtained in the time control group. Epinephrine increased LVP to that of the time control (106 +/- 3) group only in the BDM + Bim group (106 +/- 3) after hypothermia, and CF increases with adenosine, 5-hydroxytryptamine, and nitroprusside were similar to that of the time control group only in the BDM + Bim group after hypothermia. All of the effects of Bim were reversed by Glib. These results indicate that Bim, given with BDM, effectively preserves myocardial function and metabolism as well as inotropic and vasodilatory reserve during long-term hypothermic preservation as if the 1-day hypothermic state had not been instituted. Because the beneficial effects of Bim are blocked by Glib, the protective effect of Bim likely results from maintained KATP channel opening. Treatment with exogenous KATP openers may prove useful in preserving cardiac function in the transplanted heart.

Animals↗
PubMed1996

KATP channels mediate late preconditioning against infarction produced by monophosphoryl lipid A.

The cardioprotective effect of myocardial preconditioning (PC) to reduce infarct size has been shown to last approximately 90 min (early PC), and then a second window of protection (SWOP or late PC) appears 24 h later. Although much work has been done to characterize early PC, little has been done to investigate potential mediators of SWOP. To that end, we have used monophosphoryl lipid A (MLA), a nontoxic endotoxin derivative, to produce SWOP and have examined the role of ATP-sensitive potassium (KATP) channels in mediating its cardioprotection. Adult mongrel dogs were given MLA (3, 10, or 35 micrograms/kg i.v.) 24 h before a 60-min left anterior descending coronary artery occlusion and 3 h of reperfusion. After reperfusion, the hearts were stained for myocardial infarction with triphenyltetrazolium. MLA produced a dose-dependent reduction in infarct size that was associated with an enhanced shortening of the monophasic action potential duration during early ischemia. To further examine the role of KATP channels, animals were treated with MLA (35 micrograms/kg) and 24 h later were administered either glibenclamide (0.3 mg/kg i.v.) or 5-hydroxydecanoate (7.5 mg/kg intracoronary over 20 min), two structurally distinct KATP-channel antagonists. Both glibenclamide and 5-hydroxydecanoate abolished the cardioprotection produced by MLA. These results demonstrate that the cardioprotective effect of late PC produced by MLA is dependent on functional KATP channels and is the first study to suggest that late PC may be the result of an increased KATP current during ischemia.

Action Potentials↗
PubMed1996

Morphine mimics the cardioprotective effect of ischemic preconditioning via a glibenclamide-sensitive mechanism in the rat heart.

Previous results from our laboratory have suggested that opioid receptors are involved in ischemic preconditioning (PC) in rat heart. Furthermore, other investigators have suggested that mu- and delta-opioid receptors mediate analgesia and hypoxic cerebral vasodilatation via opening of ATP-sensitive K+ (KATP) channels. Thus, the purpose of the present study was to test the hypothesis that activation of opioid receptors mimics the cardioprotective effect of ischemic PC and that this effect is produced by activation of KATP channels in the rat heart. Anesthetized open-chest Wistar rats were subjected to six different protocols. All groups were subjected to 30 minutes of occlusion and 2 hours of reperfusion. Ischemic PC was elicited by three 5-minute occlusion periods interspersed with 5 minutes of reperfusion. Similarly, morphine-induced PC was elicited by three 5-minute drug infusions (100 micrograms/kg i.v. ) interspersed with 5-minute drug-free periods before the prolonged 30-minute occlusion. Infarct size (IS) as a percentage of the area at risk (AAR) was determined by triphenyltetrazolium staining. Ischemic PC and morphine infusions resulted in similar reductions in IS/AAR from 56 +/- 5% to 11 +/- 3% and 12 +/- 5%, respectively (P < .05). Administration of glibenclamide (0.3 mg/kg i.v.), a KATP channel antagonist, or naloxone (3 mg/kg i.v.), a nonselective opioid receptor antagonist, both blocked the cardioprotective effects of morphine. These results indicate that opioid receptor stimulation results in a reduction in infarct size similar to that produced by ischemic PC. The effect of morphine is most likely mediated via an opioid receptor-KATP channel-linked mechanism in the rat heart, since glibenclamide abolished its protection.

Animals↗
PubMed1996

PD 81,723, an allosteric enhancer of the A1 adenosine receptor, lowers the threshold for ischemic preconditioning in dogs.

PD 81,723 (PD) acts allosterically to increase agonist binding to A1 adenosine receptors and to enhance functional A1 receptor-mediated responses in the heart and other tissues. To determine if PD lowers the threshold for ischemic preconditioning (PC), pentobarbital-anesthetized dogs were subjected to 60 minutes of left anterior descending coronary artery (LAD) occlusion and 3 hours of reperfusion. Ischemic PC was produced by either 2.5 or 5 minutes of LAD occlusion 10 minutes before the 60-minute occlusion. PD (100 micrograms/kg total dose, 5 to 50 mumol/L in coronary arterial blood) or vehicle was infused intracoronarily for 17.5 minutes before the 60-minute occlusion period in non-PC dogs or in dogs preconditioned with 2.5 minutes of ischemia. Myocardial infarct size was determined by triphenyltetrazolium staining and expressed as a percentage of the area at risk. Compared with the control group (26.3 +/- 3.6%, mean +/- SEM), infarct size was not significantly affected by 2.5 minutes of PC alone (23.4 +/- 4.2%) or by PD alone (26.5 +/- 1.7%) but was decreased by PD + PC (14.6 +/- 1.7%, P < .05) or by a longer period (5 minutes) of PC alone (12.5 +/- 3.3%). The intravenous administration of the selective antagonist of A1 adenosine receptors, 8-cyclopentyl-1,3-dipropylxanthine (1 mg/kg), or the ATP-sensitive K+ channel blocker, glibenclamide (0.3 mg/kg), for 15 minutes before PD + PC blocked the protection (23.6 +/- 2.3% or 25.9 +/- 3.3%, respectively). None of the compounds studied affected systemic hemodynamics, collateral blood flow, or AAR. To determine which subtypes of canine adenosine receptors were affected by 10 mumol/L PD, radioligand binding studies were conducted using membranes derived from COS-7 cells expressing recombinant canine receptors and agonist radioligands. PD enhanced the binding of [125I]N6-4-amino-3-iodobenzyladenosine (125I-ABA) to A1 receptors by increasing the t1/2 for dissociation by 2.18-fold, but PD had no effect on the dissociation kinetics of 125I-ABA from A3 receptors or [125I]-[2-(4-amino-3-iodo-phenyl)ethylamino] adenosine from A2A receptors. Glibenclamide at concentrations up to 10 mumol/L had no effect on the binding of radioligands to recombinant canine A1, A2A, or A3 receptors. These data suggest that PD reduces the amount of time required for ischemia to produce preconditioning by enhancing adenosine binding to its A1 receptor. Glibenclamide prevents the protection afforded by A1 receptor activation by a mechanism not involving adenosine receptor blockade.

Allosteric Regulation↗
PubMed1995

Bimakalim, an ATP-sensitive potassium channel opener, mimics the effects of ischemic preconditioning to reduce infarct size, adenosine release, and neutrophil function in dogs.

BACKGROUND: The primary goal of the present study was to determine whether the infarct size-reducing effect of preconditioning is associated with an increase in adenosine release from the ischemic myocardium during a prolonged occlusion period or the subsequent reperfusion period and by a decrease in neutrophil infiltration. A second objective was to determine whether bimakalim, a KATP channel opener, mimics the effects of ischemic preconditioning. METHODS AND RESULTS: Barbital-anesthetized open-chest dogs were subjected to 60 minutes of left anterior descending coronary artery (LAD) occlusion followed by 3 hours of reperfusion. In the preconditioning group, 5 minutes of LAD occlusion followed by 10 minutes of reperfusion was elicited before the 60-minute occlusion period. In two other groups, bimakalim 1 microgram/kg bolus followed by a 0.05 micrograms.kg-1.min-1 infusion or an equivalent volume of saline was administered intravenously 15 minutes before occlusion and continued until the time of reperfusion. In a final group, bimakalim was administered 10 minutes before reperfusion and continued until the end of the experiment. To measure the release of adenosine from the ischemic region, coronary venous blood samples were collected at various times during ischemia and after reperfusion, and the concentration of adenosine was measured. Myocardial infarct size was determined by triphenyl tetrazolium chloride; transmural myocardial blood flow, by radioactive microspheres. Transmural myeloperoxidase (MPO) activity, an index of neutrophil infiltration in the area at risk, was also measured. Preconditioning produced a marked reduction in infarct size (9.8 +/- 3.0% versus 28.6 +/- 5.2% in the control group, mean +/- SEM); adenosine release at 5, 10, 15, and 30 minutes of the 3-hour reperfusion period; and transmural MPO activity in the risk area. Similarly, pretreatment with bimakalim resulted in reductions in infarct size, adenosine release, and transmural MPO activity to an extent almost identical to that of preconditioning. When bimakalim was administered 10 minutes before reperfusion, the drug also produced a significant reduction in infarct size and transmural MPO activity; however, no significant reduction in coronary venous adenosine concentrations was observed. There were no significant differences in collateral blood flow between groups. CONCLUSIONS: These results indicate that myocardial preconditioning in the canine heart produced by a short period of ischemia or a KATP channel opener is not mediated by an increase in adenosine release, as measured by coronary venous adenosine concentrations, during 60 minutes of occlusion or the initial 30 minutes of reperfusion. A significant reduction in transmural MPO activity in the ischemic area also appears to result from KATP channel activation and may play a role, at least in part, in the reduction in infarct size observed, particularly when a KATP channel opener is administered just before reperfusion.

Adenosine↗