beta-Adrenoceptor blocking drugs and isoprenaline: central effects on cardiovascular parameters.

1. The central hypotensive activity of (+)- and (-)-propranolol (100 microgram), pindolol (100 microgram) and isoprenaline (1 and 4 microgram) injected intracerebroventricularly (i.c.v.) was studied in rats anaesthetized with urethane and chloralose. Blood pressure, cardiac output and heart rate were measured; systolic stroke volume and peripheral vascular resistance were calculated. 2. (+)- and (-)-Propranolol and pindolol induced a fall of blood pressure but (+)-propranolol was less active. The heart rate was reduced more by (-)-propranolol than by (+)-propranolol or (-)-pindolol. The decrease of systolic stroke volume was greater for (-)-propranolol and pindolol than for (+)-propranolol. Peripheral vascular resistance was reduced to the same level but with different time courses, (-)-propranolol having a longer effect than (+)-propranolol and pindolol. 3. Isoprenaline induced a hypotensive effect, while cardiac output and heart rate increased; the systolic stroke volume remained stable but peripheral vascular resistance was significantly decreased. 4. These results suggest that different central regulatory centres are involved in the control of cardiac function and peripheral vascular tone.

Presynaptic beta-adrenoceptors in rat atria: evidence for the presence of stereoselective beta 1-adrenoceptors.

1. Presynaptic beta-adrenoceptor activity was studied in rat isolated atria, previously loaded with [3H]-noradrenaline. The stimulation-induced release of 3H transmitter was measured in the presence of cocaine, and adrenaline was used as a facilitatory beta-adrenoceptor agonist. 2. Adrenaline (0.1 and 2 nM) increased, by about 50%, the evoked efflux of tritium. With phenoxybenzamine present, the same activity was shown with 10 nM adrenaline. 3. The beta 2-selective adrenoceptor blocking drugs: IPS 339 and ICI 118 551 caused a concentration-dependent decrease in the activity of adrenaline. Cardioselective beta-blocking drugs: acebutolol, beta-xolol, nebivolol and its isomers (R 67 138 and R 67 145) also reduced dose-dependently the agonistic action of adrenaline. The order of potency for nebivolol and its isomers was R 67 138 greater than nebivolol greater than R 67 145. The activity of pindolol was not concentration-dependent. The inhibitory effect of acebutolol was also observed in the presence of blockade of alpha-adrenoceptors. 4. The postsynaptic beta-adrenoceptor blocking activity of nebivolol and its isomers was studied in pithed rats. They reduced isoprenaline-induced tachycardia without altering hypotensive responses. The order of potency was: R 67 138 greater than nebivolol greater than R 67 145. 5. It is concluded that in rat isolated atria, presynaptic beta 2- and beta 1-adrenoceptors coexist and that facilitatory beta 1-adrenoceptors are stereospecific.

Biochemical characterization of the mechanisms involved in the 5-hydroxytryptamine-induced increase in rat atrial rate.

Several possible mechanisms for 5-hydroxytryptamine (5-HT)-induced tachycardia in rat have been suggested: an activation of 5-HT1C or 5-HT2 receptors, an indirect sympathomimetic effect or a mechanism independent of 5-HT2 receptor stimulation. The aim of this study was to investigate the involvement of these mechanisms in the 5-HT-induced increase in rat atrial rate using biochemical methods. Indeed, the 5-HT1C and 5-HT2 receptors are linked to phosphoinositide hydrolysis and the noradrenaline (NA) released by 5-HT can stimulate the beta 1-adrenergic receptors linked to adenylate cyclase stimulation. The effect of varying concentrations of 5-HT on inositol phospholipid hydrolysis and adenylate cyclase activity of the rat isolated atria were measured. 5-HT (2 microM) did not modify total inositol phosphate (IP) production, while 5-HT 10 and 50 microM increased it 2-fold. The 5-HT2 antagonist ketanserin (1 microM) abolished IP accumulation induced by 5-HT microM), which indicates that this accumulation is 5-HT2 and not 5-HT1C receptor-mediated. Moreover, cyclic AMP (cAMP) formation was enhanced by 5-HT (5, 10, 20 and 50 microM). When atria were incubated 10 min with the beta-adrenergic receptor antagonist nadolol (1 microM), the increase in the cAMP level induced by 5-HT, whatever its concentration (10, 20 or 50 microM), was inhibited. Treating rats with reserpine (2.5 mg/kg, i.p., 48 and 24 hr before experimentation), which caused NA depletion in the heart, seemed to reduce the stimulating effect of 5-HT 10 and 50 microM on adenylate cyclase activity. Thus, the 5-HT-induced increase in cAMP is indirectly due to the activation of the beta-adrenergic receptors by the NA released by 5-HT. It is concluded that 5-HT stimulates both phosphoinositide turnover and adenylate cyclase activity in the rat isolated atria by activation of 5-HT2 receptors and by an indirect sympathomimetic effect.

Chronic subcutaneous treatment with acebutolol: haemodynamic effects and metabolism in spontaneously hypertensive rats.

Chronic administration of acebutolol (15 mg kg-1 s.c. three times a week for five weeks, then 30 mg kg-1 for three weeks) did not lower blood pressure in 17 and 33 weeks-old spontaneously hypertensive rats (SHR). At the end of this treatment, the plasma concentrations of acebutolol and diacetolol were measured by HPLC. After 24 h, acebutolol was absent from plasma while diacetolol was lower after chronic treatment than after acute administration. Twenty-four hours after the last injection of acebutolol, both isoprenaline-induced tachycardia and vasodilatation were reduced. The vasomotor agents, noradrenaline, bradykinin and angiotensin, exhibited the same activity in control and treated SHR. These findings suggest that the lack of antihypertensive effect of acebutolol in SHR may be the result of a decrease in diacetolol formation together with blockade of beta 2 vascular receptors.

Cardiovascular effects of intracerebroventricular injections of (+/-)-nebivolol and its enantiomers--a comparison with those of metoprolol in the rat.

The cardiovascular effects of (+/-)-nebivolol, a potent beta 1-adrenoceptor antagonist, and its enantiomers, (+)-nebivolol (SRRR) and (-)-nebivolol (RSSS) in normotensive anaesthetized rats, have been investigated using metoprolol as a reference substance. The drugs decreased blood pressure and heart rate immediately after i.c.v. injection. These effects paralleled the beta-blocking potencies ((+)- greater than (+/-)-greater than (-)-nebivolol). Metoprolol induced a weaker hypotension than (+/-)-nebivolol, and a long-lasting reduction in stroke volume. As reported after i.v. administration, (+/-)-nebivolol and isomers by the i.c.v. route decreased peripheral vascular resistance following i.c.v. administration while metoprolol increased it. These effects are centrally mediated since cardiovascular responses to isoprenaline i.v. remained unchanged.

5-Hydroxytryptamine and beta-adrenoceptors in rat isolated atria.

The positive chronotropic effect of a high concentration of 5-hydroxytryptamine (5-HT) in rat isolated atria results mainly from a tyramine-like mechanism and is linked to an increase in cAMP production by an indirect stimulation of beta-adrenoceptors. Using this preparation, we have compared the action of tyramine and 5-HT. The tyramine (0.15 microM)-induced increase in atrial rate was suppressed by atenolol (a beta 1-blocking drug) and by nadolol (a beta 1 beta 2-blocker), while the positive chronotropic effect of 5-HT was reduced by atenolol and suppressed by nadolol. The 5-HT-induced elevation in cAMP was unchanged in the presence of atenolol and abolished by nadolol. The involvement of beta 2-adrenoceptors in the effects of 5-HT could result from competition between 5-HT and noradrenaline at the beta 1-adrenoceptors that results in a fixation of noradrenaline on beta 2-adrenoceptors.

Acute cardiovascular activity of beta-adrenoceptor blocking drugs in awake or anaesthetized normotensive and hypertensive rats.

The effects of intravenous dl-propranolol (5 mg/kg), 1- and d-alprenolol (5 mg/kg), on blood pressure, heart rate, cardiac output and total peripheral resistance were investigated in normotensive and spontaneously, renal or neurogenic hypertensive rats. The animals were either awake or anaesthetized with urethane, chloralose or pentobarbital. Anaesthesia may induce hemodynamic changes in hypertensive rats; thus urethane decreased peripheral vascular resistance in spontaneously hypertensive rats. In anaesthetized rats, dl-propranolol, 1- and d-alprenolol caused significant falls in blood pressure, heart rate and cardiac output. The hypotensive activity was related to the initial blood pressure. Peripheral vascular resistance remained unchanged except in spontaneously hypertensive, chloralose anaesthetized rats. 1-Alprenolol had less cardiac effects than dl-propranolol. The decrease of blood pressure was shorter and/or smaller after d-alprenolol than after 1-alprenolol. In normotensive and renal hypertensive awake rats, dl-propranolol induced a short fall of blood pressure, due to a decrease of heart rate and cardiac output. These effects, except bradycardia, were not observed in spontaneously hypertensive rats. These results suggest that differences in the initial vascular state, in the hypertensive models and in the anaesthetics used can explain the variability of the responses to beta-adrenergic blocking drugs. Moreover, beta-blocking as well as quinidine-like activities are involved in the cardiovascular responses to dl-propranolol, d-alprenolol and 1-alprenolol studied here.

[Autonomic cardiovascular regulation and permanent catheterization in normotensive conscious (WKY) and spontaneously hypertensive (SHR) rats]. / Régulation cardiovasculaire autonome et cathétérisme permanent chez le rat éveillé normotendu (WKY) et spontanément hypertendu (SHR).

In cardiovascular research, methods of indwelling catheterism have been frequently described. In the present work, we used normotensive and SHR rats to compare carotid catheterism to left ventricular catheterism, this last method being proposed by some investigators for cardiac index measurement. Our results of plasma catecholamines and autonomous nervous system activity show that ventricular catheterism results in an important disturb in cardiovascular regulation and question its validity to study cardiac function.

Tissue distribution of acebutolol in mature normotensive and stroke-prone spontaneously hypertensive rats.

Tissue distribution of acebutolol was studied in 33-week-old normotensive (WKY) and Okamoto stroke-prone (SHR-SP) rats, 30 min after an i.v. administration, by using 14C-acebutolol. Plasma level of acebutolol was higher in WKY than in SHR-SP. Aorta, kidney, liver and muscle radioactivity/plasma radioactivity ratios were higher in SHR-SP than in WKY. The brain/plasma radioactivity ratio was very low and similar in the two groups. The drug distribution was the same in the two groups except in medulla + corpus trapezoides where drug concentration was greater in SHR-SP. These results, compared with previous ones, show an age-related evolution in pathological state in SHR-SP. They point out a specific concentration of the beta-blocking drug in a defined part of the brain, namely medulla + corpus trapezoides.

Study on the mechanism of 5-HT-induced tachycardia in the pithed rat.

1. Intravenous infusion of serotonin (5-HT) (2.5, 5, 10 and 20 micrograms/kg/min) in pithed rats induced a dose-dependent sustained tachycardia. 2. Pretreatment by phentolamine or diltiazem did not modify the chronotropic response to 5-HT. In contrast, atenolol antagonized this tachycardia and the 5-HT antagonists methysergide, ketanserin and MDL 72222 reduced it. 3. The 5-HT-induced tachycardia was abolished by desipramine and was not affected by fluvoxamine, a specific 5-HT uptake inhibitor. Surrenalectomy did not change the response to 5-HT but catecholamine depletion by reserpine markedly inhibited it. 4. Infusion of 5-HT increased the ratio of noradrenaline (NA) in the heart to NA in plasma, from 1.70 in control group to 2.76 in treated group (P less than 0.05). Desipramine inhibited this effect. 5. It was concluded that the tachycardia induced by an infusion of 5-HT in pithed rat results from a complex mechanism involving mainly the release of NA from the cardiac sympathetic nerves and a less important direct 5-HT2 mechanism.

Uptake of 5-hydroxytryptamine in rat isolated atria.

1. The mode of 5-hydroxytryptamine (5-HT) uptake by the rat isolated atria was studied and compared to noradrenaline (NA) uptake. 2. Rat isolated atria were incubated with 14C-5-HT (46 microM) or 3H-NA 0.4 microM). After washing, the radioactivity fixed in atria was counted and the total NA, 5-HT and 5-hydroxyindol-3-acetic acid (5-HIAA) atria contents were measured by HPLC. 3. 14C-5-HT uptake was reduced in atria from 6-hydroxydopamine (100 mg/kg, i.p., 48 hr before experiments) or reserpine (2.5 mg/kg, i.p., 24 and 48 hr before experiments) pretreated rats. 4. The incubation of atria with 5-HT (50 microM) at the same time as 3H-NA reduced the 3H-NA value fixed. 5. Addition of desipramine (1 microM) or hydrocortisone (150 microM) before the incubation of atria with 14C-5-HT was without any effect on 14C-5-HT uptake. In contrast, fluvoxamine (1 microM) or indalpine (5 microM) caused a slight inhibition. 6. These data indicate that 5-HT is taken into the NA storage vesicles within the atria sympathetic nerves. This uptake does not use the NA carrier and involves partly the 5-HT carrier. An extraneuronal accumulation was noticed and a part of it is intracellular.

[Role of 5-HT2 receptors in the positive chronotropic action of serotonin on the isolated atria in rats]. / Rôle des récepteurs 5-HT2 dans l'action chronotrope positive de la sérotonine sur les oreillettes isolées de rat.

5-hydroxytryptamine (5-HT), from the concentration of 5 microM to 50 microM has a positive chronotropic activity on the rat isolated atria. The tyramine-like indirect sympathomimetic effect is the main mechanism of the 5-HT action at high concentration (50 microM). In this study, we have demonstrated that 5-HT2 receptor stimulation is also involved in the 5-HT-induced increase in atrial rate. Thus, when the tyramine-like effect was inhibited by reserpine, which causes a noradrenaline depletion of atria, the positive chronotropic effect of 5-HT (50 microM) was reduced but not abolished. The remaining response to 5-HT was completely inhibited by the 5-HT2 antagonist ketanserin (1 microM). Moreover, the increase in atrial rate induced by 5-HT, at the lower concentration of 5 microM, was fully abolished by this antagonist.

Peripheral and central cardiovascular effects of ketanserin in conscious normotensive rats.

The aim of the present study was to evaluate the effects of low doses of ketanserin, close to therapeutic ones, in conscious normotensive rats and to compare the different thresholds of 5-HT2- and alpha-antagonist properties and central sympatho-inhibitory activity. Male Sprague-Dawley rats (250-300 g) were used. Fourty-eight hr before experimentation, indwelling venous and arterial catheters were placed in each rat under light anaesthesia, and exteriorized through the interscapular skin. Drugs injected by i.v. route were ketanserin (50, 100, 200 micrograms/kg), serotonin (50 micrograms/kg), noradrenaline (1 microgram/kg), angiotensin (0.5 microgram/kg), nitroglycerin (200 micrograms/kg) and histamine (100 micrograms/kg). Ketanserin induced a short-lasting reduction in blood pressure and did not modify heart rate. At the dose of 50 micrograms/kg, this compound fully inhibited the pressor effect of serotonin, without modifying its hypotensive responses; after 200 micrograms/kg, ketanserin reduced by 50% the pressor effect of noradrenaline. Comparison of antagonistic activities on phenylephrine and noradrenaline pressor responses showed a selective blockade of alpha 1-adrenoceptors by 100 micrograms/kg ketanserin. In contrast, the angiotensin hypertensive response was unchanged. Reflex bradycardia after noradrenaline and angiotensin was reduced by 100 micrograms/kg ketanserin, whereas nitroglycerin and histamine effects were unaffected. In conclusion, administration of increasing low doses of ketanserin showed that blockade of 5-HT2-receptors appears in first, then a central sympatho-inhibitory effect and a selective alpha 1-adrenolytic activity occur. In addition, baroreceptor sensitivity seems not to be affected.

Cardiovascular effects of drugs acting on the autonomous nervous system, in awake, normotensive and hypertensive rats.

A study has been carried out in awake normotensive (WKY), spontaneously hypertensive (SHR) and renal hypertensive (RHR) rats in order to compare the reactivity of the autonomous nervous system using different indwelling catheterizations. In all cases, a jugular catheter was implanted for i.v. injections. The arterial cannula was implanted in the femoral artery (group 1) or in the right carotid artery (group 2), where the catheter was routed over 20 mm towards the heart, reaching the aortic arch. The plasma catecholamines levels were lower in the RHR than in the WKY or SHR, and whatever the strain, lower in group 2 than in group 1. In the WKY and the RHR, the mean blood pressure was higher in group 2 than in group 1. The tachycardia induced by isoprenaline and the pressor response to noradrenaline were higher in the SHR than in the WKY and in the RHR. By contrast, the reflex bradycardia was lower. Furthermore, the procedure of catheterization could modify the cardiovascular responses induced by adrenergic and cholinergic agents. These results demonstrate that indwelling catheterizations cannot be performed safely in the rat when studying effects on cardiac performance or vascular reactivity.

Central action of beta-adrenoceptor antagonists on blood pressure after acute administration in rats.

The effect of an i.v. administration of some beta-adrenergic blocking drugs on blood pressure has been investigated in rats after blood-brain barrier (BBB) opening. Practolol and atenolol which do not penetrate the BBB, induced an immediate hypotensive effect after BBB breakdown by intracarotid (i.c.) injection of cetrimonium. In 39 week-old spontaneously hypertensive rats, practolol (15 mg/kg i.v.) and atenolol (3 mg/kg i.v.) induced a large drop in blood pressure while only a slight decrease was shown in normotensive ones. Likewise, acebutolol induced a significantly greater hypotension after BBB damage. On the other hand, the effects of dl-propranolol (5 mg/kg), quinidine (2.5 mg/kg) and isoproterenol (3 micrograms/kg) on blood pressure were not modified by pretreatment with cetrimonium i.c., while the hypotension induced by d-propranolol (5 mg/kg) was shortened. These results indicate that beta-adrenergic blocking agents with a low degree of lipophily can induce a hypotensive effect when their penetration into brain is largely enhanced after BBB opening either by prolonged hypertension or by cetrimonium. This effect is only dependent on their action on beta-adrenoreceptors; membrane stabilizing effect and intrinsic sympathomimetic activity do not seem to be involved.

[In vitro metabolism of 4-benzylisoquinolines, analogs of papaverine]. / Contribution à l'étude du métabolisme in vitro de benzyl-4 isoquinoleines, analogues de la papavérine.

A comparative study was made on sliced Rat liver of the in vitro disappearance of papaverine, PV 2 6,7-dimethoxy-4-(parachlorobenzyl)isoquinoline and its mono and di isopropoxy derivatives. Data show that papaverine and PV 2 are equally sensitive to oxygenases, although 7 mono and 6,7 di-isopropoxy derivatives are much less so, being more bulky and undergo in enzymic O-dealkylation less easily. The data also show that PV 2 and 6-isopropoxy desmethoxy PV 2 disappear at the same rate, more readily than the 7 isopropoxy isomer. This confirms the in vivo results previously reported. The antispasmodic in vitro activity of these compounds is reported.

cGMP release in rat mesenteric arterioles and in conduit mesenteric artery.

1. Relaxing factors were studied in two perfused preparations of the same vascular area in the rat: resistance mesenteric arterioles and conduit mesenteric artery. 2. In both preparations, an acetylcholine (ACh) infusion inhibited noradrenaline (NA) vasoconstrictor effects but at a ten-times greater concentration in conduit artery than in resistance arterioles. 3. Endothelium destruction with hypotonic Krebs solution did not change basal perfusion pressure, but increased NA responses and suppressed ACh inhibitory effects in arterioles and arteries. Likewise, L-NAME abolished the ACh effect in mesenteric arterioles but only reduced it in mesenteric artery. 4. Basal release of cyclic GMP was significantly greater in mesenteric artery than in resistance arterioles. By contrast, ACh-induced cGMP release was higher in mesenteric arterioles. Endothelium removal did not change basal release of cGMP in mesenteric arterioles but reduced it in mesenteric artery. 5. These results suggest that in basal conditions several relaxing factors are present in higher concentrations in conduit mesenteric artery than in resistance mesenteric arterioles. However, although it releases higher basal amount of cGMP, this vessel has a reduced role in vascular control than do smaller arteries.

Acute haemodynamic effects and tissue distribution of acebutolol in normotensive and spontaneously hypertensive rats.

Acute i.v. administration of 15 mg/kg acebutolol in normotensive (WKY), Okamoto (SHR) and Okamoto stroke-prone (SHR-SP) awake rats resulted in weak effects on blood pressure and in bradycardia more marked in SHR-SP. Thirty minutes after i.v. administration, lung and renal uptake of [14C]acebutolol was reduced in SHR compared to other rats. Muscle uptake was higher in SHR and blood concentration was higher in SHR-SP. Brain levels were low and similar in all rats. Plasma protein binding was identical in all rats. These results are discussed according to the characteristics of the three strains studied.

Modulation of noradrenergic transmission in the rat isolated portal vein: role of prejunctional alpha 2-adrenoceptors and beta-adrenoceptors.

1. The effect of several adrenoceptor agonists and antagonists on the spontaneous and stimulus-evoked release of [3H]noradrenaline was studied in rat isolated portal vein. 2. Yohimbine (10(-6)M) increased the stimulus-evoked [3H]noradrenaline efflux. Adrenaline alone (3 x 10(-6)M) was without effect, while it increased the resting efflux when added together with yohimbine. 3. Propranolol alone was without effect on the release of [3H]noradrenaline. When added (2 x 10(-6)M) at the same time as yohimbine, it reduced the stimulation-induced 3H efflux. When added before adrenaline and yohimbine, propranolol (10(-5)M) reduced both spontaneous and stimulus-evoked release of [3H]noradrenaline. 4. The effect of several beta-blocking drugs was measured on the enhancing effect of yohimbine on the stimulation-evoked 3H efflux. The beta 1-adrenoceptor blocking drugs: atenolol (5 x 10(-6) and 10(-5) M), metoprolol (5 x 10(-6) and 10(-5) M), like the more selective bisoprolol (2 x 10(-6) and 4 x 10(-6) M) inhibited yohimbine activity. Likewise, propranolol (2 x 10(-6) and 5 x 10(-6)M) and the beta 2-adrenoceptor blocker ICI 118551 exhibited an antagonistic effect. 5. These results indicate the possibility for noradrenaline to activate presynaptic beta-adrenoceptors in rat portal vein. They show an interaction between the presynpatic alpha 2- and beta-adrenoceptor mediated systems in the release of noradrenaline. They suggest the presence and the activity of facilitatory beta 1-adrenoceptors.