Refresher course for teaching cardiovascular physiology.
This report presents highlights of a refresher course presented at Experimental Biology '99 on Saturday, April 17, 1999, in Washington, District of Columbia.
Curriculum↗
Biomedical subjects
F L Belloni
Publications and source records attributed to F L Belloni.
Teaching the principles of hemodynamics.
Knowledge of hemodynamic principles is crucial to an understanding of cardiovascular physiology. This topic can be effectively taught by discussing simple physical principles and basic algebraic equations. A variety of examples from everyday observations can be used to illustrate the physical principles underlying the flow of blood through the circulation, thereby giving the student an experiential feel for the topic in addition to an understanding of theory. Moreover, opportunities abound for showing how each hemodynamic principle can explain one or another functional feature of the cardiovascular system or a cardiovascular pathophysiological state. Thus hemodynamics can be used as an organizational thread to tie together other aspects of cardiovascular physiology.
Blood Pressure↗
Preconditioning with ischemia or adenosine protects skeletal muscle from ischemic tissue reperfusion injury.
Prolonged tissue ischemia and subsequent reperfusion results in significant tissue injury due to the ischemic-reperfusion (IR) syndrome. Ischemic preconditioning (IPC) or adenosine (ADO) pretreatment are known to protect IR injury in cardiac muscle. Our aim was to determine whether IPC or ADO pretreatment attenuates and protects against ischemic tissue reperfusion injury in skeletal muscle. Rats were anesthetized and global hindlimb ischemia was induced by 60 min of suprarenal aortic clamping followed by 30 min of reperfusion period. The degree of skeletal muscle dysfunction was determined by decreases in maximum contractile force, and adenosine triphosphate (ATP) and creatine phosphate (CP) levels of extensor digitorum longus (EDL) muscle. The distal tendon of the EDL was attached to a force transducer for maximum isometric force measurement. Samples were taken from the EDL for measurement of ATP and CP levels. The following were protective protocols prior to the IR challenge: (1) four consecutive 5-min periods of ischemia separated by 5-min reperfusion periods (PC/I) or (2) i.v. adenosine infusion (350 microg/kg/min x 10 min, PC/A). Our data suggest that pretreatment with brief periods of ischemia or systemic ADO infusion attenuates ischemic tissue reperfusion injury in skeletal muscle. [Table: see text]
Adenosine Diphosphate↗
Role of nitric oxide in hypoxic coronary vasodilatation in isolated perfused guinea pig heart.
To test the hypothesis that nitric oxide (NO) mediates hypoxic coronary dilatation in situ, isolated guinea pig hearts were perfused at constant pressure (Langendorff technique) with physiological salt solution. Switching from a control perfusate (95% O2-5% CO2) to one equilibrated with a lower O2 tension (20% O2) induced a large, but submaximal and reproducible, coronary dilatation. The NO synthase inhibitor NG-nitro-L-arginine (L-NNA) diminished baseline flow (3.67 +/- 0.24 vs. control 5.11 +/- 0.42 ml.min-1 x g-1; P < 0.05) and selectively blocked the coronary flow response to acetylcholine without reducing the response to papaverine. L-NNA reduced the absolute increase in coronary flow during hypoxia by 27 +/- 2% (delta flow = 5.83 +/- 0.49 vs. control delta flow = 8.04 +/- 0.74 ml.min-1 x g-1; P < 0.05). Hypoxic coronary dilatation was unaffected by infusion of the thromboxane mimetic U-46619, which decreased baseline coronary flow to the same extent as L-NNA. Prior addition of indomethacin did not alter the attenuating effect of L-NNA. Hypoxic coronary dilatation during constant flow perfusion at 14.7 +/- 0.28 ml/min was reduced by 65 +/- 5% after L-NNA. Therefore, the NO component of the response was not a consequence of the reduced baseline flow observed in the presence of L-NNA, did not depend on prostaglandin synthesis, and was not secondary to increased flow or intravascular shear stress. We conclude that hypoxic coronary vasodilatation in isolated guinea pig hearts is partially mediated by NO.
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
Differential sensitization of cardiac actions of adenosine in rats after chronic theophylline treatment.
To determine the effect of chronic adenosine receptor blockade on atrial responsiveness, we administered theophylline to rats in their drinking water (0.6 mg/ml) for 2 wk. Inotropic and chronotropic responses to the adenosine receptor agonists N6-cyclopentyladenosine (CPA) and 5'-(N-ethylcarboxamido)-adenosine (NECA) were then measured in isolated atria from treated and control animals. The indirect (antiadrenergic) actions of CPA and NECA on force and rate, measured during beta-adrenergic receptor stimulation by isoproterenol, were markedly sensitized (2- to 10-fold reductions in the agonist concentration needed to obtain a half-maximal response) after theophylline. The sensitization was homologous because inotropic and chronotropic responses to carbachol were not affected by theophylline. The direct negative inotropic and chronotropic actions of CPA and NECA, measured without concomitant beta-adrenergic stimulation, were not sensitized after theophylline. The number of atrial A1-receptors, measured by antagonist radioligand binding (maximum specific binding at saturation), was increased by 22% in theophylline-treated rats [66.2 +/- 3.4 vs. 54.3 +/- 1.9 (control) fmol/mg protein, P < 0.05]. Competition binding indicated that the fraction of coupled (high-affinity) receptors was unchanged. The number of ventricular A1-receptors was increased to a similar extent without any change in coupling. Thus chronic dietary theophylline upregulated cardiac A1-adenosine receptors without changing coupling state or affinity and sensitized rat atria to the indirect, antiadrenergic, inhibitory inotropic and chronotropic actions of adenosine receptor agonists.
Adenosine↗
Cardiac desensitization to adenosine analogues after prolonged R-PIA infusion in vivo.
To determine the effects of chronic in vivo stimulation of adenosine receptors, R-(-)-N6-(2-phenylisopropyl)adenosine (R-PIA), a selective A1 receptor agonist, was administered to rats as a continuous 7-day infusion (200 nmol/h). Inotropic and chronotropic responses of isolated atria to adenosine receptor agonists were markedly desensitized compared with the responses of atria from age-matched control animals. Carbachol's negative chronotropic effect was also attenuated, indicating a heterologous mode of desensitization. Antagonist radioligand binding assays indicated a 52% reduction in A1 adenosine receptor maximum binding, and competition binding assays revealed a significant loss of G protein-coupled high-affinity A1 receptors in atria from R-PIA-treated rats. Inhibitory G proteins (Gi) were significantly reduced, as quantified by immunoblot analysis, with no change in the amount of stimulatory G proteins. Ventricular membranes from R-PIA rats showed loss of Gi and uncoupling of A1 receptors, without a significant change in A1 receptor density. Thus chronic R-PIA infusion desensitized rat atrial muscle to the effects of adenosine receptor agonists via several regulatory adaptations, including downregulation of A1 adenosine receptors, uncoupling of A1 receptors from their associated G proteins, and loss of Gi proteins.
2-Chloroadenosine↗
Mechanisms of coronary vasodilatation produced by ATP in guinea-pig isolated perfused heart.
1. Isolated hearts of guinea-pigs were perfused in vitro with a physiological salt solution via a retrograde aortic cannulation (Langendorff preparation) at constant perfusion pressure. Bolus intra-arterial injections of various vasodilator drugs were made and the coronary flow responses were measured with an electromagnetic flow probe placed in the arterial inflow circuit. Inhibitory drugs were infused intra-arterially. 2. Nitro-L-arginine (NLA; 500 microM), an NO synthesis inhibitor, decreased coronary baseline flow by 16 +/- 0.8%, converted acetylcholine-induced coronary vasodilatation to vasoconstriction and had no effect on coronary flow responses to adenosine or papaverine. Sodium nitroprusside-induced responses were enhanced during NLA infusion by 46 +/- 11%. 3. Adenosine 5'-triphosphate (ATP) increased coronary flow but coronary flow responses to ATP were not altered by infusion of NLA. 4. ATP-induced coronary dilatation was not significantly attenuated by infusion of the adenosine receptor antagonist XAC, (xanthine amine congener; 2 microM), whereas XAC decreased coronary flow responses to adenosine by 75% +/- 5%. 5. ATP-induced coronary flow responses were reduced by only 31 +/- 4% during indomethacin infusion (2.8 microM) whereas indomethacin completely eliminated the initial vasoconstriction phase and greatly attenuated the peak flow and duration of the later vasodilatation phase seen in response to arachidonic acid (0.75 nmol). Indomethacin had no effect on vasodilatations produced by adenosine or prostaglandin I2. 6. These results indicate that ATP-induced coronary dilatation in the isolated, perfused heart of the guinea-pig is not dependent upon NO production or upon degradation of ATP to adenosine. The coronary dilator action of ATP may be partially dependent (approximately 30%) upon the production of vasodilator prostaglandins.
Adenosine Triphosphate↗
Vasodilative and anti-adrenergic effects of adenosine in diabetic rat hearts.
To determine the vasodilative and negative inotropic effects of adenosine in hearts of diabetic rats, isolated hearts, perfused at constant perfusion pressure (Langendorff technique), were prepared from age-matched control Wistar rats and rats made diabetic 10 weeks prior to study by a single injection of streptozotocin (65 mg.kg-1, i.p.). Adenosine and nitroprusside each increased coronary inflow when administered either as bolus injections or as infusions. Coronary flow responses to nitroprusside were unchanged in diabetic hearts. Coronary flow responses of diabetic hearts to adenosine injections were unchanged, but responses to adenosine infusions tended to be larger than in normal hearts. Diabetes had no significant effect on the EC50 for either vasodilator. Adenosine inhibited the inotropic effect of isoproterenol (enhanced left ventricular (LV) pressure (P) and LV dP/dtmax) in normal hearts, independently of its vasodilative action. This negative inotropic action of adenosine appeared equally strong in diabetic hearts. We conclude that adenosine's coronary vasodilative and anti-beta-adrenergic, negative inotropic effects in the rat heart were not diminished after 10 weeks of streptozotocin-induced diabetes mellitus. Thus, earlier reports of diminished adenosine dilative efficacy in experimental diabetes may have been unique to those particular models.
Adenosine↗
Adenosine causes bradycardia in pacing-induced cardiac failure.
BACKGROUND: In normal, conscious dogs, systemic injection of adenosine causes arterial hypotension and a baroreceptor reflex tachycardia mediated in part by withdrawal of vagal tone from the sinoatrial node. After vagal section or muscarinic receptor blockade, however, adenosine injection causes bradycardia via a direct sinoatrial node inhibition. Because cardiac failure is marked by a loss of vagal tone, we hypothesized that adenosine injection in dogs with failing hearts would reduce heart rate. METHODS AND RESULTS: Mongrel dogs were instrumented with indwelling catheters, manometers, and ventricular pacing electrodes. After the dogs had recovered from the surgery, the ventricles were paced continuously at 210 beats per minute for 3 weeks, followed by pacing at 240 beats per minute for an additional week. This regimen caused mild ventricular and more striking atrial hypertrophy and a gradual onset of physiological and clinical signs of congestive heart failure. Adenosine injections that caused large tachycardias before the pacing regimen began caused progressively smaller increments in heart rate during the first 2 weeks of pacing. After 3 and 4 weeks, adenosine injections caused overt reductions in heart rate despite the concomitant arterial depressor response. CONCLUSIONS: We conclude that the loss of vagal tone associated with the development of cardiac failure unmasks the direct negative chronotropic effect of exogenous adenosine on the sinoatrial node.
Adenosine↗
Glibenclamide attenuates adenosine-induced bradycardia and coronary vasodilatation.
The effects of the ATP-sensitive K(+)-channel blocker glibenclamide on the cardiovascular responses to adenosine in dogs were determined. Adenosine (0.01-20 mumol/kg iv) caused coronary vasodilatation, arterial hypotension, and bradycardia in dogs with either combined beta-adrenergic and muscarinic receptor blockade or with bilateral cervical vagotomy plus beta-adrenergic receptor blockade. The 50% effective dose for adenosine-induced coronary dilatation was increased from 0.13 +/- 0.04 mumol/kg in the control state to 1.1 +/- 0.5 mumol/kg after 2 mg/kg of glibenclamide (P less than 0.001). Adenosine at 5 mumol/kg reduced heart rate by 19 +/- 5% from a baseline of 158 +/- 6 beats/min in five anesthetized dogs. After glibenclamide (10 mg/kg), this dose of adenosine failed to cause a significant change in heart rate. The arterial hypotensive effects of adenosine were also attenuated by glibenclamide. Thus glibenclamide inhibited adenosine-induced bradycardia, hypotension, and coronary dilatation. On the other hand, glibenclamide did not affect the reductions in heart rate caused by vagus nerve stimulation. The mechanism of this adenosine antagonism is not known but, in the case of bradycardia, it does not appear to involve any of the steps shared in common by both adenosine-induced and vagal responses of the sinoatrial node.
Adenosine↗
Mechanism of the apparent parasympathetic inhibition of adenosine induced heart rate slowing in the dog.
The inhibitory action of intravenously administered adenosine on the sinoatrial (SA) node is not expressed in the conscious dog in the presence of normal vagal tone. After pharmacological or surgical parasympathetic blockade, however, adenosine exerts a powerful negative chronotropic effect. In order to determine the reason why this action of adenosine is blocked by the intact parasympathetic nervous system, we measured the chronotropic effects of adenosine while applying a constant cholinergic stimulus to the SA node. In conscious dogs during a systemic infusion of acetylcholine at a rate sufficient by itself to inhibit the SA node, intravenous adenosine injections caused further dose dependent reductions in heart rate. In anaesthetised, vagotomised dogs, intravenous adenosine caused similar negative chronotropic effects with or without concomitant electrical stimulation of the vagus nerve. As an example, 5 mumol.kg-1 adenosine reduced heart rate by 22 (SEM 4)% from a baseline heart rate of 172(10) beats.min-1; when heart rate was lowered to 66(1) beats.min-1 by electrical vagal stimulation, this dose of adenosine reduced heart rate by 36(8)%. Propranolol had no effect on these responses. We conclude that there is no direct cholinergic inhibition of the negative chronotropic action of adenosine on the canine SA node but rather that the inhibitory action of systemically administered adenosine on the SA node is simply masked by the withdrawal of vagal tone in response to the arterial hypotension resulting from this mode of adenosine administration.
Acetylcholine↗
An unusual receptor mediates adenosine-induced SA nodal bradycardia in dogs.
To characterize the receptor mediating the negative chronotropic effect of adenosine in dogs, experiments were performed on conscious dogs with chronically implanted cardiovascular instrumentation. Autonomic blockade was used to eliminate any reflex influences on heart rate. Intravenous bolus injections of various adenosine analogues caused dose-dependent, aminophylline-blockable reductions in heart rate with a potency order of 5'-(N-ethylcarboxyamido)-adenosine (NECA)-78:2-chloroadenosine-17:adenosine-1. Dipyridamole enhanced the potency of adenosine to equal that of 2-chloroadenosine. Moderately selective A1-receptor agonists N6-(L-2-phenylisopropyl)-adenosine (R-PIA) and N6-cyclohexyladenosine and an A2-selective agonist 2-phenylaminoadenosine (200 nmol/kg) had no negative chronotropic effect in the conscious dog. Adenosine and its analogues, including R-PIA, caused coronary vasodilatation at smaller doses than were required to slow the heart rate. The selective A1-adenosine receptor blocker xanthine amine congener (XAC) antagonized the negative chronotropic action of adenosine but did so nonselectively, as the coronary vasodilative and negative chronotropic actions of adenosine were antagonized equally well. The spontaneous contraction rate of isolated perfused dog right atrial preparations, which included the sinoatrial node, was reduced by intrasinoatrial node artery infusions of adenosine analogues with a potency ratio of NECA-100:adenosine-15:N6-cyclopentyladenosine-2.3:R-PIA-1. We conclude that the adenosine receptor mediating the negative chronotropic action of adenosine in the dog does not display the pharmacological characteristics of either typical A1- or A2-adenosine receptors. Instead, either a novel adenosine receptor or an A1-receptor with unusual agonist and antagonist binding properties appears to exist in the dog's sinoatrial node.
2-Chloroadenosine↗
Effects of alkylxanthines and calcium antagonists on adenosine uptake by cultured rabbit coronary microvascular endothelium.
Adenosine uptake by cultured rabbit coronary microvascular endothelial cells was studied. Radiolabeled [2-3H]-adenosine, present initially in the extracellular space at 10(-6) mol/l, was incorporated into the cell cultures at a steady rate during 30 s-3 h incubations. Incorporated 3H was found mostly (83%) in adenine nucleotides. Incorporation of [3H]-adenosine was attenuated by an adenosine deaminase inhibitor (EHNA) but only at adenosine concentrations of 10(-5) mol/l or higher. Adenosine transport inhibitors (dipyridamole, nitrobenzylthioinosine) attenuated 3H incorporation. Adenosine uptake was also diminished by certain structural analogues of adenosine (e.g., 2-chloroadenosine), by several alkylxanthine drugs (theophylline, isobutylmethylxanthine, enprofylline and 8-phenyltheophylline), and by certain calcium antagonists (verapamil, nifedipine and trifluoperazine). The mechanisms of actions of these agents on adenosine uptake do not appear to be related to phosphodiesterase inhibition, adenosine receptor antagonism or calcium antagonism. The effects of varying adenosine metabolism may contribute to the pharmacologic actions of these agents.
1-Methyl-3-isobutylxanthine↗
Effects of adenosine on the heart rate in conscious animals: interaction between adenosine and parasympathetic mechanisms at the sinoatrial node.
Explore the source record for details and available documents.
Adenosine↗
Uptake and release of adenosine by cultured rat aortic smooth muscle.
We wanted to determine whether CO2, H+ and K+ affect the adenosine metabolism of vascular smooth muscle in a way that could account for the effects of these substances on vascular reactivity and their ability to modulate adenosine-induced vascular relaxation. Accordingly, 1-week-old cultures of rat aortic smooth muscle were incubated in phosphate-buffered saline with various [K+]'s and pH's and aerated in an incubation chamber with gases containing various proportions of CO2. Uptake was measured as 14C incorporation into cellular constituents during exposure to 2 microM [14C]adenosine. Release was measured as net extracellular adenosine accumulation. Uptake of adenosine was not significantly affected by any of the experimental maneuvers, except that it was greatly attenuated by dipyridamole (10(-5) and 10(-4) M) and transiently enhanced by the low CO2 levels. Adenosine release, however, was depressed by lowering atmospheric CO2 (0% vs 5%) and also by normocapnic acidosis (pH 6.8 vs pH 7.4). We conclude that vascular smooth muscle in culture releases adenosine at a rate that might have vasoactive significance in vivo. Furthermore, some of the vascular actions of CO2 and H+, but not those of K+, may be partially explained by their effects on vascular smooth muscle's adenosine metabolism.
Adenosine↗
The mechanism of adenosine release from hypoxic rat liver cells.
Uptake of [14C]-adenosine into freshly dispersed rat hepatocytes was inhibited 44% by dipyridamole (50 microM) and 60% by nitrobenzylthioinosine (NBTI, 20 microM). The results are consistent with the known ability of these drugs to inhibit adenosine transport in other cell types. The nucleotide analogue, alpha, beta-methylene adenosine diphosphate (AOPCP, 50 microM), inhibited by 84% the degradation of exogenous 5' AMP that occurred rapidly when this substrate alone was presented to isolated hepatocytes. This confirms the ecto-5'-nucleotidase inhibitory properties of this analogue in isolated hepatocytes. During hypoxic incubation, isolated hepatocytes released adenosine, which accumulated in the extracellular volume. Dipyridamole and NBTI each markedly attenuated this extracellular adenosine accumulation. In contrast, AOPCP had no inhibitory effect on net hypoxic adenosine release. It is concluded that hypoxic rat hepatocytes produce adenosine intracellularly and that this adenosine is released via facilitated diffusion to the extracellular space, based on the inhibition observed with the transport inhibitors. The plasma membrane enzyme ecto-5'-nucleotidase does not appear to participate in hypoxic adenosine release from these cells as indicated by the lack of effect of the nucleotidase inhibitor, AOPCP.
Adenosine↗
Apparent reduction in baroreflex sensitivity to adenosine in conscious dogs.
Relative effects of equihypotensive doses (-35 mmHg) of adenosine (5.0 mumol/kg) and nitroglycerin (25 micrograms/kg) on heart rate and, therefore, baroreflex sensitivity were studied in conscious dogs. Nitroglycerin increased heart rate 133 +/- 24% from 78 +/- 5.5 beats/min, whereas adenosine increased heart rate only 79 +/- 16% from 78 +/- 5.2 beats/min (P less than 0.01). Injection of nitroglycerin during combined beta-adrenergic and muscarinic receptor blockades caused arterial pressure to fall 38 +/- 3.4% from 107 +/- 3.2 mmHg without any significant change in heart rate (3.8 +/- 3.8 from 162 +/- 9.2 beats/min). During combined beta-adrenergic and muscarinic receptor blockades adenosine also reduced arterial pressure 45 +/- 2.7% from 106 +/- 2.9 mmHg but unexpectedly reduced heart rate as well by 37 +/- 1.7% from 160 +/- 9.7 beats/min. This bradycardia reflected an effect on the sinoatrial (SA) node rather than an induction of heart block, since the R-R interval increased by 70 +/- 7.8% from 371 +/- 20 ms (P less than 0.01), while the P-R interval increased only 13 +/- 2.3% from 97 +/- 7.2 ms (P less than 0.05) with no electrocardiographic evidence of nonconducted beats. Arterial plasma adenosine levels were 43 +/- 5 nmol/ml at this time. Adenosine also caused bradycardia during muscarinic blockade alone (-43 +/- 3.4% from 201 +/- 6.4 beats/min) and following bilateral vagal section (-33 +/- 1.9% from 151 +/- 5.9 beats/min). In summary, adenosine appears to alter normal baroreflex function in the conscious dog by reducing the tachycardia that normally follows a fall in systemic arterial pressure.(ABSTRACT TRUNCATED AT 250 WORDS)
Adenosine↗
Superoxide anion selectively attenuates catecholamine-induced contractile tension in isolated rabbit aorta.
Xanthine oxidase-derived oxygen metabolites caused a selective loss of norepinephrine-induced contractile tension in rings and helical strips from rabbit aorta. Phenylephrine-induced tension was not affected. The relaxation was selectively and completely blocked by superoxide dismutase but not by catalase. Isoproterenol-induced relaxation was also reversed by xanthine oxidase-derived oxygen metabolites. These observations are consistent with the chemical reaction of superoxide anion with catecholamines and suggest that the reaction may have significance at physiological concentrations of norepinephrine. The time course of the effects of superoxide generation on contractile tension was consistent with the properties of the chemical reaction (measured spectrophotometrically) and with the dependence of tone on norepinephrine concentration. These results indicate that superoxide anion, in situations at which submicromolar concentrations of this reduced oxygen metabolite are present, will selectively oxidize catecholamines, which may attenuate local adrenergic regulation.
Animals↗