Pharmacological modulation of b-adrenoceptors as a new cardioprotective strategy for therapy of myocardial dysfunction induced by ischemia and reperfusion

Abstract Purpose: To evaluate the cardioprotective response of the pharmacological modulation of β-adrenergic receptors (β-AR) in animal model of cardiac ischemia and reperfusion (CIR), in spontaneously hypertensive (SHR) and normotensive (NWR) rats. Methods: CIR was induced by the occlusion of left anterior descendent coronary artery (10 min) and reperfusion (75 min). The SHR was treated with β-AR antagonist atenolol (AT, 10 mg/kg, IV) 5 min before CIR, and NWR were treated with β-AR agonist isoproterenol (ISO, 0.5 mg/kg, IV) 5 min before CIR. Results: The treatment with AT increased the incidence of VA, AVB and LET in SHR, suggesting that spontaneous cardioprotection in hypertensive animals was abolished by blockade of β-AR. In contrast, the treatment with ISO significantly reduced the incidence of ventricular arrhythmia, atrioventricular blockade and lethality in NWR (30%, 20% and 20%, respectively), suggesting that the activation of β-AR stimulate cardioprotection in normotensive animals. Serum CK-MB were higher in SHR/CIR and NWR/CIR compared to respective SHAM group (not altered by treatment with AT or ISO). Conclusion: The pharmacological modulation of β-AR could be a new cardioprotective strategy for the therapy of myocardial dysfunctions induced by CIR related to cardiac surgery and cardiovascular diseases.

Participation of hepatic α/β-adrenoceptors and AT1 receptors in glucose release and portal hypertensive response induced by adrenaline or angiotensin II

It has been previously demonstrated that the hemodynamic effect induced by angiotensin II (AII) in the liver was completely abolished by losartan while glucose release was partially affected by losartan. Angiotensin II type 1 (AT1) and adrenergic (∝1- and β-) receptors (AR) belong to the G-proteins superfamily, which signaling promote glycogen breakdown and glucose release. Interactive relationship between AR and AT1-R was shown after blockade of these receptors with specific antagonists. The isolated perfused rat liver was used to study hemodynamic and metabolic responses induced by AII and adrenaline (Adr) in the presence of AT1 (losartan) and ∝1-AR and β-AR antagonists (prazosin and propranolol). All antagonists diminished the hemodynamic response induced by Adr. Losartan abolished hemodynamic response induced by AII, and AR antagonists had no effect when used alone. When combined, the antagonists caused a decrease in the hemodynamic response. The metabolic response induced by Adr was mainly mediated by ∝1-AR. A significant decrease in the hemodynamic response induced by Adr caused by losartan confirmed the participation of AT1-R. The metabolic response induced by AII was impaired by propranolol, indicating the participation of β-AR. When both ARs were blocked, the hemodynamic and metabolic responses were impaired in a cumulative effect. These results suggested that both ARs might be responsible for AII effects. This possible cross-talk between β-AR and AT1-R signaling in the hepatocytes has yet to be investigated and should be considered in the design of specific drugs.

Interação entre as vias de sinalização de receptores serotoninérgicos e Β-adrenérgicos em artéria femoral de ratos / Interaction between serotoninergic-and β-adrenergic receptors signaling pathways in rat femoral artery / Interacción entre las vías de señalización de receptores serotoninérgicos y β-adrenégicos en la arteria femoral de ratones

FUNDAMENTO: A doença coronária tem sido amplamente estudada em pesquisas cardiovasculares. No entanto, pacientes com doença arterial periférica (DAP) têm piores resultados em comparação àqueles com doença arterial coronariana. Portanto, os estudos farmacológicos com artéria femoral são altamente relevantes para a melhor compreensão das respostas clínicas e fisiopatológicas da DAP. OBJETIVO: Avaliar as propriedades farmacológicas dos agentes contráteis e relaxantes na artéria femoral de ratos. MÉTODOS: As curvas de resposta de concentração à fenilefrina contrátil (FC) e à serotonina (5-HT) e os agentes relaxantes isoproterenol (ISO) e forskolina foram obtidos na artéria femoral de ratos isolada. Para as respostas ao relaxamento, os tecidos foram contraídos com FC ou 5-HT. RESULTADOS: A potência de classificação na artéria femoral foi de 5-HT > FC para as respostas contráteis. Em tecidos contraídos com 5-HT, as respostas de relaxamento ao isoproterenol foram praticamente abolidas em comparação aos tecidos contraídos com FC. A forskolina, um estimulante da adenilil ciclase, restaurou parcialmente a resposta de relaxamento ao ISO em tecidos contraídos com 5-HT. CONCLUSÃO: Ocorre uma interação entre as vias de sinalização dos receptores β-adrenérgicos e serotoninérgicos na artéria femoral. Além disso, esta pesquisa fornece um novo modelo para estudar as vias de sinalização serotoninérgicas em condições normais e patológicas que podem ajudar a compreender os resultados clínicos na DAP.

BACKGROUND: Coronary heart disease has been widely studied in cardiovascular research. However, patients with peripheral artery disease (PAD) have worst outcomes compared to those with coronary artery disease. Therefore, pharmacological studies using femoral artery are highly relevant for a better understanding of the pathophysiologic responses of the PAD. OBJECTIVE: The aim of this study was to evaluate the pharmacologic properties of the contractile and relaxing agents in rat femoral artery. METHODS: Concentration response curves to the contractile phenylephrine (PE) and serotonin (5-HT) and the relaxing agents isoproterenol (ISO) and forskolin were obtained in isolated rat femoral artery. For relaxing responses, tissues were precontracted with PE or 5-HT. RESULTS: The order rank potency in femoral artery was 5-HT > PE for contractile responses. In tissues precontracted with 5-HT, relaxing responses to isoproterenol was virtually abolished as compared to PE-contracted tissues. Forskolin, a stimulant of adenylyl cyclase, partially restored the relaxing response to ISO in 5-HT-precontracted tissues. CONCLUSION: An interaction between β-adrenergic- and serotoninergic- receptors signaling pathway occurs in femoral artery. Moreover, this study provides a new model to study serotoninergic signaling pathway under normal and pathological conditions which can help understanding clinical outcomes in the PAD.

FUNDAMENTO: La enfermedad coronaria ha sido ampliamente estudiada en las investigaciones cardiovasculares. Sin embargo, los pacientes con enfermedad arterial periférica (EAP), tienen los peores resultados en comparación con aquellos con la enfermedad arterial coronaria. Por tanto, los estudios farmacológicos con la arteria femoral son extremadamente importantes para obtener una mejor comprensión de las respuestas clínicas y fisiopatológicas de la EAP. OBJETIVO: Evaluar las propiedades farmacológicas de los agentes contráctiles y relajantes en la arteria femoral de los ratones. MÉTODOS: Las curvas de concentración-respuesta a los agentes conctráctiles fenilefrina (FE) y a la serotonina (5-HT) y los agentes relajantes isoproterenol (ISO) y forskolina, se obtuvieron en la arteria femoral de ratones ya aislada. Para las respuestas a la relajación, los tejidos fueron contraídos con FE o 5-HT. RESULTADOS: La potencia de clasificación en la arteria femoral fue de 5-HT > FE para las respuestas contráctiles. En los tejidos contraídos con 5-HT, las respuestas de relajación al isoproterenol fueron prácticamente eliminadas en comparación con los tejidos contraídos con FE. La forskolina, un estimulante de la adenilil ciclasa, restauró parcialmente la respuesta de relajación al ISO en los tejidos contraídos con 5-HT. CONCLUSIÓN: Ocurre una interacción entre las vías de señalización de los receptores β-adrenérgicos y serotoninérgicos en la arteria femoral. Además, esa investigación suministra un nuevo modelo para estudiar las vías de señalización serotoninérgicas en condiciones normales y patológicas que puedan ayudar a comprender los resultados clínicos en la EAP.

Zinc and water intake in rats: investigation of adrenergic and opiatergic central mechanisms

We have demonstrated that central administration of zinc in minute amounts induces a significant antidipsogenic action in dehydrated rats as well as in rats under central cholinergic and angiotensinergic stimulation. Here we show that acute third ventricle injections of zinc also block water intake induced by central ß-adrenergic stimulation in Wistar rats (190-250 g). Central inhibition of opioid pathways by naloxone reverses the zinc-induced antidipsogenic effect in dehydrated rats. After 120 min, rats receiving third ventricle injections of isoproterenol (160 nmol/rat) exhibited a significant increase in water intake (5.78 ± 0.54 ml/100 g body weight) compared to saline-treated controls (0.15 ± 0.07 ml/100 g body weight). Pretreatment with zinc (3.0, 30.0 and 300.0 pmol/rat, 45 min before isoproterenol injection) blocked water intake in a dose-dependent way. At the highest dose employed a complete blockade was demonstrable (0.54 ± 0.2 ml/100 g body weight). After 120 min, control (NaAc-treated) dehydrated rats, as expected, exhibited a high water intake (7.36 ± 0.39 ml/100 g body weight). Central administration of zinc blocked this response (2.5 ± 0.77 ml/100 g body weight). Naloxone pretreatment (82.5 nmol/rat, 30 min before zinc administration) reverted the water intake to the high levels observed in zinc-free dehydrated animals (7.04 ± 0.56 ml/100 g body weight). These data indicate that zinc is able to block water intake induced by central ß-adrenergic stimulation and that zinc-induced blockade of water intake in dehydrated rats may be, at least in part, due to stimulation of central opioid peptides

Comparative study of beta-adrenoceptor blocking efficacy of two formulations of esmolol in vivo and in vitro.

The objective of the present study was to compare the cardiovascular beta-blocking activity of two different formulations of esmolol. Spontaneously beating guinea-pig isolated atria and the heart rate and blood pressure of anaesthetized cat were employed in the study to compare the beta-blocking efficacy of the two formulations of esmolol using isoprenaline as an agonist. In guinea-pig isolated atria the standard esmolol formulation (Brevibloc) reduced basal atrial rate more significantly than the indigenously formulated esmolol (test formulation). Both the formulations produced similar parallel rightward shift of cumulative concentration response curves of isoprenaline with closely comparable pA2 values. In anaesthetized cats, only indigenous esmolol formulation significantly decreased basal heart rate. Both the formulations did not modify the basal blood pressure and isoprenaline-induced fall in blood pressure, despite significantly blocking isoprenaline-induced tachycardia. It is suggested that both the formulations produced similar degree of beta-1 adrenoceptor blocking activity.

Experimental Chagas' disease: effects of propranolol upon cardiac beta-receptors

Chagas' disease is an important cause of heart disfunction in Latin America. Previous works from our laboratory reproducing experimental Chagas' disease in mice, demonstrated that the affinity and density of cardiac beta-adrenergic receptors were altered during the acute, indeterminate and chronic phase in Albino Swiss mice inoculated with Trypanosoma cruzi. Keeping in mind that Propranolol is a beta-blocking agent that binds in the same receptors'site, which we have described as altered along T. cruzi infection. The present study was performed to determine if a beta-blocker treatment could prevent cardiac beta-receptors'disorders provoked by T. cruzi infection. Two different doses of Propranolol (9 and 40 mg/Kg/day) were injected in the mice during 3 days; then they were infected with 7 x 10(4) parasites/mouse and propranolol was continued daily for one week. The results showed that the concentrations of propranolol used did not protect the beta-receptors'sites by administration of each doses.

Beta-adrenoreceptor blockade attenuates heat-induced tachycardia,but not the tolerance to the stress.

Ten healthy males (age 34 +/- 3 yr 9 SE) underwent 40 min of heat exposure (WD 39.7.C) after 2 hours of ingesting 120 mg of Propranolol (Inderal; ICI), or a placebo, in a random manner, the exposures being about a week apart. That there was no placebo effect was ensured by giving a control run (no medication). In the placebo trials, the end-experiment heart rate had increased by 52%, while after propranolol the increase was only 43%. Regression analysis showed that with the placebo, the HR increased by 22 beats/min/o rise in core (aural) temperature, while with propranolol, the rise (14 beats/min) was significantly lower (P < 0.02). The various heat strain indices viz the Craig's Index, the Body heat storage (Kilocals/m2/hr), and the effective heat storage were also similar for both the treatments. We conclude that beta-adrenoreceptor activity plays a significant role in producing tachycardia of heat exposure in humans, but blocking this activity with propranolol does not affect tolerance to heat stress.

Immunopharmacological studies on Picrorhiza kurroa Royle-ex-Benth. Part IV: Cellular mechanisms of anti-inflammatory action.

In this work abroagation of anti-inflammatory effect of Picrorhiza kurroa extract (PK) by beta-adrenergic blockade was confirmed, which suggests alteration in cell-surface biology by PK treatment. Blockade of protein synthesis by cycloheximide pretreatment reduced PK effect, suggesting protein mediation. Metabolic inhibitor dinitrophenol inhibited inflammatory cedema equally in control and PK treated animals, and masking of PK effect was concluded. Discriminations of anti-inflammatory mechanism(s) of PK and the latter two cytotoxic agents was inferred from these observations and from existing knowledge. Selective PK influence on membrane linked activation events in inflammatory effector cells could be the basis of anti-inflammatory and perhaps other biological activities reported with the herb.

Immunopharmacological studies on Picrorhiza kurroa Royle-ex-Benth. Part III: Adrenergic mechanisms of anti-inflammatory action.

Nature of adrenergic mechanisms contributing to anti-inflammatory effect of Picrorhiza kurroa suggested from earlier studies was explored in Wistar albino rats. Water soluble fraction of alcoholic extract of rhizomes (PK) potentiated castor oil-catharsis on oral administration but direct subplanter PK-injections failed to exhibit any local irritancy and oedema. Propranolol pretreatment counteracted while phentolamine enhanced anti-inflammatory effect of PK in carrageenin-induced inflammation. 6-hydroxy-dopamine pretreatment antagonised the said PK-effect and in such animals both ephedrine and isoprenaline augmented anti-inflammatory effect of PK, the former interaction being more conspicuous. PK-treatment of rats did not influence adrenaline uptake by lung slices in vitro. The results suggest that a non-neural augmentation of beta-adrenoceptor function or consequent cellular events mediates the anti-inflammatory effect of PK.

Peripheral action of angiotensin II.

The effects of angiotensin II were studied on isolated atrial preparations of nonreserpinised and reserpinised rabbits, before and after treating the preparations by propranolol. Peripheral action of angiotensin was cardioaccelerator via direct stimulation of beta-receptors of the atria in isolated atrial preparations.

Effect of propranolol and phenoxybenzamine on muscle strength and physical endurance, and on exercise-induced changes in muscle glycogen and blood lactic acid levels in rats.

Propranolol and phenoxybenzamine (PBZ) had no major effect on swimming-endurance performance and skeletal muscle pulling-strength in rats. Propranolol, like exercise itself diminished the resting skeletal muscle glycogen (SMG) content, but the drug did not lower the resting blood lactic acid (BLA) levels. Propranolol significantly antagonised the 25-min exercise-induced BLA elevation, indicating that lactacidaemia is possibly a beta-receptor response. However, propranolol did not exhibit an overall beneficial effect on swimming endurance. PBZ had a negligible effect on maximal swimming time (MST) and had no major effects on the SMG content and BLA levels.

Adrenergic receptors in the hypothalamus concerned with control of arterial blood pressure: elicited by electrical and chemical stimulation.

The study was made with the help of a chemitrode placed in various areas of the hypothalamus by the stereotaxic technique, for electrical and chemical stimulation (noradrenaline or isoprenaline) before and after microinjection of respective blockers (phenoxybenzamine or practalol). The results indicated the presence of both alpha and beta-adrenoreceptors in the anterior and dorsomedial hypothalamus producing a depressor response, and, the presence of alpha adrenoreceptors in the posterior and lateral hypothalamus producing a pressor response.

Reduced beta adrenergic responsiveness in isolated rabbit atria during hypothermia.

Beta adrenergic agonist isoprenaline (8x10(-6)M) produced a 500% increase in inotropic and 90% increase in the chronotropic responses of isolated rabbit atria at 37 degrees C. On cooling the atria to 23 degrees C, these responses were significantly reduced to 87% and 30% respectively. Similar results were obtained with adrenaline, but isoprenaline was more potent. The positive chronotropic and inotropic responses to isoprenaline were effectively blocked by propranolol and practolol at 37 degrees C whereas at 23 degrees C these beta blockers were unable to block even minor positive responses obtained by isoprenaline at this temperature. On the contrary at 23 degrees C, phenylephrine (alpha adrenergic agonist) produced marked positive chronotropic and inotropic effects indicating enhancement of alpha adrenoceptor activity at lower temperatures. This also suggests that reduced beta receptor activity at lower temperature is not due to a generalised depression of adrenoceptors as a result of hypothermia. Rewarming of atria to 37 degrees C restored the beta adrenoceptor responsiveness to previous level. It appears that ambient temperature is important im maintaining normal beta adrenergic activity of the atria.