[Adipocytokines: implications in the prognosis and drug treatment of cardiovascular diseases]. / Adipocitocinas: implicaciones en el pronóstico y tratamiento farmacológico de patología cardiovascular.

Adipocytokines, fat tissue derived factors with regulatory properties, are involved in the pathophysiology of atheromatous and metabolic illnesses such as: ischemic heart disease, insulin resistance, obesity, dyslipidemia and diabetes mellitus. Enlargement of visceral adipose tissue depots determines a worse evolution for those complaints. Drugs as angiotensin converting enzyme inhibitors (ACEI), thiazolidinediones (glitazones) or angiotensin-II receptor antagonists, generally associated with the adequate hypolipidemic (statins, fibrates) or antiobesity (orlistat, sibutramine, rimonabant) medication, would increase those adipocytokines with anti-inflammatory and insulin-sensitizing properties (i.e. adiponectin or visfatin), while reducing pro-inflammatory and thrombogenic cytokines (as leptin, tumor necrosis factor [TNF]-alpha, plasminogen activator inhibitor 1 [PAI-1]). Thus, these pharmacologic therapeutic approaches would have a beneficial effect in order to diminish morbidity-mortality and improve the prognosis of patients with said diseases, all of them related to high cardiovascular risk.

Effects of verapamil and elgodipine on isoprenaline-induced metabolic responses in rabbits.

Verapamil (0.17 microg kg(-1) min(-1) intravenous, i.v.) but not elgodipine (35 ng kg(-1) min(-1)) modestly enhanced the weak blood glucose increase induced by the i.v. infusion of isoprenaline (0.3 microg kg(-1) min(-1)) in conscious rabbits. However, elgodipine but not verapamil suppressed the increase in circulating insulin evoked by the agonist. Both drugs enhanced the rise in plasma lactate mediated by isoprenaline but only elgodipine potentiated the lipolytic effect of the agonist. In isolated islets elgodipine (10(-6) M) blocked forskolin (10(-6) M)-induced insulin release. However, in rabbit adipocytes elgodipine potentiated both glycerol release and cAMP accumulation induced by isoprenaline (10(-8)-10(-6) M). Excess K(+) (40-60 mM) did not alter basal lipolysis or the response to isoprenaline in either rabbit or mouse adipocytes. Therefore, Ca2+ influx through L-type Ca2+ channels does not seem to play a significant role in the lipolytic effect of isoprenaline. Metabolic alterations found with Ca2+ channel antagonists were of minor intensity and probably devoid of pathological implications.

Role of alpha2-adrenoceptors on the hyperglycaemic and insulin secretory effects derived from alpha1- and beta-adrenoceptor stimulation in the rabbit.

1. In conscious fasted rabbits the insulin secretory response induced by the intravenous infusion of the alpha1-adrenoceptor agonist, amidephrine (10 microg kg(-1) min(-1)) was blocked by the simultaneous administration of clonidine (2 microg kg(-1) min(-1) i.v.). 2. The excitatory effect of amidephrine (10 microg kg(-1) min(-1)) on insulin secretion was similarly suppressed by the concomitant infusion of the selective alpha2-adrenoceptor agonist UK14304 (1 microg kg(-1) min(-1)). Both, the increase in blood glucose and the inhibition of insulin secretion found with UK14304 when infused alone were antagonized in rabbits previously treated with the very selective alpha2-adrenoceptor antagonist 2-methoxyidazoxan (1.5 microg kg(-1) min(-1)). 3. The combined administration of amidephrine (3 microg kg(-1) min(-1)) and isoprenaline (0.3 microg kg(-1) min(-1)) evoked a potentiated increase in insulin plasma levels in the face of a weak hyperglycaemia, an established reduction in blood pressure and tachycardia. 4. The potentiated insulin secretory response derived from alpha1- and beta-adrenoceptor stimulation was blunted by clonidine administration. In its presence a sustained hyperglycaemic response was found. 5. The increase in plasma lactate levels resulting from dual adrenoceptor stimulation (amidephrine: 10 microg kg(-1) min(-1) + salbutamol: 0.3 microg kg(-1) min(-1)) was smaller than the expected should addition or potentiation occurred. 6. Our results point to a possible physiological role played by alpha2-adrenoceptors on insulin secretion, since their stimulation by the endogenous catecholamines could lead to inhibition of insulin release, masking any potentiated response that otherwise should have appeared from alpha1- and beta-adrenoceptor stimulation.

Comparative effects of calcium channel blockers and indomethacin on insulin secretion and blood pressure increase derived from alpha 1-adrenoceptor stimulation in the rabbit.

1. In conscious, fasted rabbits the intravenous infusion of the alpha 1-adrenoceptor agonist, amidephrine (3 and 10 micrograms kg-1 min-1) induced a dose related increase in insulin plasma levels. This effect was accompanied by a minor hypo- or hyperglycaemic response, depending on the dose of agonist infused. 2. A dose related increase in mean arterial pressure and reduction in heart rate were also found after amidephrine administration. 3. The insulin secretory response to amidephrine was not prevented in rabbits previously treated with atropine (5.26 micrograms kg-1 min-1). However, in the presence of muscarinic receptor blockade the bradycardic effect of amidephrine was either suppressed or attenuated. 4. Pretreatment with the calcium channel antagonist elgodipine (35 ng kg-1 min-1) or with indomethacin (0.66 mg kg-1 min-1) clearly blocked the effect of amidephrine on insulin secretion. 5. The haemodynamic changes induced by amidephrine were preserved in the presence of either verapamil (0.17 microgram kg-1 min-1) or indomethacin, whereas the hypertensive response was antagonized by elgodipine. 6. Our results suggest that the metabolic and haemodynamic changes mediated by amidephrine are two independent effects, insulin secretion requiring the presence of extracellular calcium and the synthesis of arachidonic acid metabolites.

Adrenergic lipolysis in guinea pig is not a beta 3-adrenergic response: comparison with human adipocytes.

beta 3-Adrenoceptor agonists are potent lipolytic activators in rats, but they are only weak stimulators in human adipocytes, indicating interspecies differences in the adrenergic regulation of lipid mobilization. Like human but not rat adipocytes, guinea pig fat cells were poorly responsive to the beta 3-agonists BRL-37344, CGP-12177, SR-58611, and ICI-215001, acid metabolite of ICI-D7114. In guinea pigs, the beta 1-agonist dobutamine was more lipolytic than the beta 2-agonist procaterol. Anatomic location of fat deposits was without major influence on the beta-adrenergic responsiveness. Weak responses to beta 3-agonists were found whatever the sex or the age (from 2 days to 16 mo) of the animals. Even in the interscapular brown adipose tissue, which is well known in rats for its beta 3-adrenergic responsiveness, a blunted response to BRL-37344 was observed. The alpha 2-adrenergic antilipolytic effect and receptor number were smaller in guinea pig than in human adipocytes, but the beta-adrenergic receptor number was similar in the two species. Thus guinea pig adipocytes resemble human fat cells when their weak beta 3-adrenergic responsiveness is considered.

Effects of salbutamol and BRL 37344 on diastolic arterial blood pressure, plasma glucose and plasma lactate in rabbits.

The aim of this study was to investigate in rabbits the diastolic arterial blood pressure, plasma glucose and plasma lactate responses to salbutamol (a selective beta-2 adrenoceptor agonist) and BRL 37344 (a selective beta-3 adrenoceptor agonist) in comparison with CGP 12177 (a potent beta-1 and beta-2 adrenoceptor antagonist which also acts as a partial beta-3 agonist), isoprenaline (a non-selective beta-1, beta-2 and beta-3 adrenoceptor agonist) and adrenaline (a non-selective beta and alpha adrenoceptor agonist). All drugs were iv infused at the same dose: 0.3 microgram/kg/min (30 min). In sodium pentobarbitone (40 mg/kg)-anasthetized animals none of these compounds altered diastolic arterial blood pressure. BRL 37344 (0.1, 0.3, 1 microgram/kg/min) did not modify this parameter either. In conscious 24-h fasted rabbits, only adrenaline was able to increase plasma glucose levels. By contrast, under the same experimental conditions, salbutamol, isoprenaline and adrenaline, but not BRL 37344 or CGP 12177, induced a significant increase in plasma lactate levels. Finally, the salbutamol-mediated plasma lactate response was inhibited in the presence of clonidine (2 micrograms/kg/min, an alpha-2 adrenoceptor agonist), a drug considered to have opposite effects (stimulatory and inhibitory) on the adenylate cyclase system. In conclusion, these data suggest that only beta-2 adrenoceptor stimulation is able to increase plasma lactate levels, a response which is inhibited by alpha-2 adrenoceptor stimulation.

Coexistence of beta 2- and beta 3-adrenoceptors in plasma potassium control in conscious rabbits.

1. In conscious rabbits the intravenous infusion of adrenaline (0.3 microgram kg-1 min-1), noradrenaline (1 microgram kg-1 min-1) or isoprenaline (1.25 micrograms kg-1 min-1) caused a significant decrease in plasma potassium levels. Propranolol (9 mg kg-1, s.c.) and ICI 118551 (30 micrograms kg-1, s.c.) reversed adrenaline-induced hypokalaemia and revealed a sustained hyperkalaemia. 2. Salbutamol (0.5 microgram kg-1 min-1, i.v.), beta 2-adrenoceptor agonist, evoked a biphasic response: an initial hyperkalaemia which was followed by a hypokalaemia; a higher dose (3 micrograms kg-1 min-1, i.v.) solely induced hypokalaemia. ICI 118551 blocked the salbutamol-mediated response. 3. Noradrenaline evoked hypokalaemia was blunted completely in the presence of bupranolol (0.1 mg kg-1, s.c.), a beta 1-, beta 2- and beta 3-adrenoceptor antagonist, but not in the presence of the beta 1-adrenoceptor antagonist CGP 207 12A (1 mg kg-1, s.c.). 4. BRL 37344 (0.15 microgram kg-1 min-1, i.v.), SR 58611A (0.26 microgram kg-1 min-1, i.v.), both full beta 3-agonists, and CGP 12177 (0,25 micrograms kg-1 min-1, i.v.), a partial agonist which also acting as a non-selective beta 1- and beta 2-antagonist, induced a significant hypokalaemia. Bupranolol, but not ICI 118551 or CGP 20712A, blocked the BRL 37344-mediated hypokalaemia. 5. Ouabain (1.7 micrograms kg-1 min-1, i.v.), an inhibitor of the Na,K-pumps, inhibited both salbutamol-and BRL 37344-mediated hypokalaemia. 6. These data suggest the coexistence of beta 2- and beta 3-adrenoceptor control of extrarenal potassium disposal; moreover both beta 2 and beta 3 hypokalaemic effects would be mediated by activation of Na,K-pumps.

Effects of catecholamines on plasma potassium: the role of alpha- and beta-adrenoceptors.

The sympathetic nervous system plays an important role in the control of plasma potassium levels. Administration of adrenaline or noradrenaline evokes, in the majority of mammal species a dual response: first a short transient hyperkalaemia, followed by a maintained hypokalaemia. Alpha 1- and alpha 2-adrenoceptors mediate the initial hyperkalaemia through the activation of hepatic Ca(2+)-dependent-K(+)-channels. Stimulation of beta 1- and beta 2-adrenoceptors induces the late hypokalaemia by stimulation of skeletal muscle Na(+)-K(+)-ATPase. Beta 3-adrenoceptor stimulation may also have an effect on plasma potassium control since administration of selective beta 3-adrenoceptor agonists induces a decrease in plasma potassium. The simultaneous infusion of phenyleprine (alpha-adrenoceptor agonist) and isoprenaline (beta-adrenoceptor agonist) increases plasma potassium levels: this effect is several times larger than the algebric summation of the changes in plasma potassium when each agent is infused separately, thus suggesting potentiation. The physiological (changes in cell volume and function secondary to changes in ion fluxes) and clinical implications (pathophysiological conditions with hypo or hyperkalaemia, hyperkalaemic periodic paralysis, ventricular arrythmias) of these findings are discussed.

Functional involvement of alpha 1- and alpha 2-adrenoceptors in 86Rb efflux from liver slices and lipolysis in guinea-pig isolated adipocytes.

1. The application of an alpha 1-adrenoceptor agonist, amidephrine, to guinea-pig liver slices increases glucose release and 86Rb efflux. Since prazosin was more potent than yohimbine in inhibiting both responses, alpha 1-adrenoceptors seem to be involved in the effects evoked by the agonist. 2. Clonidine (an alpha 2-adrenoceptor agonist) at doses unable to activate liver glycogenolysis increased 86Rb release and potentiated isoprenaline in promoting 86Rb efflux. Since yohimbine antagonized clonidine in promoting 86Rb efflux, alpha 2-adrenoceptors also seem to control plasmalemmal permeability to 86Rb. 3. The liver slice responses resulting from alpha 1- and alpha 2-adrenoceptor stimulation required extracellular calcium. Calcium absence or the administration of D-600 attenuated the effects of amidephrine on glucose release and 86Rb outflow and Ca2+ excess re-established both responses. D-600 and apamin blocked clonidine-induced 86Rb efflux, suggesting that alpha 2-adrenoceptor stimulation activates calcium dependent K+ channels. 4. alpha 2-adrenoceptors do not appear to mediate antilipolytic effects in guinea-pig fat cells.

Role of Ca2+ channel blockers in insulin secretion resulting from alpha 1- and beta-adrenoceptor stimulation in the rabbit.

In conscious fasted rabbits the i.v. infusion of amidephrine (alpha 1-agonist) or isoprenaline (beta-agonist) induced an increase in plasma levels of immunoreactive insulin. The alpha 1-mediated response was suppressed in animals pretreated with calcium channel blockers (verapamil and elgodipine). The potentiated insulin secretory response in rabbits exposed to dual (alpha 1 + beta) adrenoceptor stimulation was also prevented by verapamil and elgodipine. These results suggest that extracellular calcium is required to mediate the effects of amidephrine on insulin secretion and to support potentiation.

Role of alpha-adrenoceptors in control of plasma potassium in conscious rabbits.

1. In conscious fed rabbits the intravenous infusion of amidephrine (10 micrograms kg-1 min-1), an alpha 1-adrenoceptor agonist, caused a significant increase in plasma potassium levels that was blunted by prazosin (50 micrograms kg-1, s.c.). Idazoxan failed to modify this response. 2. Clonidine (2 micrograms kg-1 min-1, i.v.), an alpha 2-adrenoceptor agonist, also evoked a hyperkalaemic response which was antagonized by idazoxan (1 microgram kg-1, s.c.), yohimbine (0.45 mg kg-1, s.c.) and prazosin (50 micrograms kg-1, s.c.). Apamin (40 micrograms kg-1, i.v. bolus) also suppressed the clonidine-mediated hyperkalaemia. 3. Verapamil (5 micrograms kg-1, s.c.) prevented both alpha 1- and alpha 2-adrenoceptor-mediated increase in plasma potassium levels. 4. It is concluded that in conscious fed rabbits both alpha 1- and alpha 2-adrenoceptor stimulation induce hyperkalaemia by activation of hepatic Ca(2+)-activated K(+)-channels.

Changes in plasma glucose and lactate evoked by alpha and beta 2-adrenoceptor stimulation in conscious fasted rabbits.

In conscious fasted rabbits, the iv infusion of salbutamol (3 micrograms/kg per min) and clonidine (2 micrograms/kg per min) induced a blood glucose increase amenable to blockade, respectively by ICI 118551 (1 micrograms/kg per min) and idazoxan (20 micrograms/kg per min). Amidephrine (10 micrograms/kg per min) and salbutamol mediated an increase in plasma lactate which was attenuated by prazosin (50 micrograms/kg, sc) and ICI 118551 respectively. Clonidine did not alter basal plasma lactate. The iv infusion of adrenaline (0.3 micrograms/kg per min) evoked an increase in plasma lactate more sensitive to blockade by ICI 118551 than by prazosin. ICI 118551 also shortened the hyperglycaemic response to adrenaline, 3-Mercaptopicolinic acid (25 mg/kg) reduced salbutamol- and adrenaline-mediated hyperglycaemia and increased at the same time the lactate/glucose ratio. Our data show that plasma lactate levels may be regulated by alpha 1- and beta 2-excitatory adrenoceptor stimulation. However, only the increase in blood lactate derived from beta 2-adrenergic stimulation seems to contribute to the overall catecholamine-mediated hyperglycaemia.

Role of alpha 1- and alpha 2-adrenoceptors in catecholamine-induced hyperglycaemia, lipolysis and insulin secretion in conscious fasted rabbits.

1. In conscious fasted rabbits an intravenous infusion of clonidine (2 micrograms kg-1 min-1) induced hyperglycaemia. The increase in blood glucose was accompanied by an inhibition of insulin secretion and basal lipolysis. 2. Yohimbine infused at a rate of 20 micrograms kg-1 min-1 suppressed clonidine-induced hyperglycaemia and blocked the inhibitory effect on insulin secretion mediated by the alpha 2-adrenoceptor agonist. 3. The intravenous infusion of amidephrine (10 micrograms kg-1 min-1) induced an increase in insulin secretion in the absence of patent hyperglycaemia. Prazosin, 0.3 mg kg-1 s.c. selectively antagonized the effect of amidephrine on insulin secretion. 4. Isoprenaline infusion (4.4 micrograms kg-1 min-1) evoked a significant increase in blood glycerol and immunoreactive insulin plasma levels. Both responses were clearly attenuated when alpha 2-adrenoceptors were simultaneously stimulated by selective (clonidine) and less selective (phenylephrine, 20 micrograms kg-1 min-1) agonists. 5. Amidephrine infusion did not induce appreciable changes in blood glycerol nor did it modify, isoprenaline-induced lipolytic response. 6. Simultaneous infusion of isoprenaline and amidephrine induced a remarkable increase in insulin secretion. 7. It is concluded that in normal fasted rabbits stimulation of alpha 2-adrenoceptors depresses basal and beta-adrenoceptor mediated lipolysis and insulin secretion. On the other hand, selective stimulation of alpha 1-adrenoceptors does not affect lipolysis but induces insulin release. Simultaneous stimulation of alpha 1- and beta-adrenoceptors potentiates the insulin secretory response.

Alpha-adrenoceptor involvement in catecholamine-induced hyperglycaemia in conscious fasted rabbits.

In conscious fasted rabbits an intravenous infusion of phenylephrine (20 micrograms kg-1 min-1) induced hyperglycaemia. The increase in blood glucose was accompanied by a modest increase in insulin secretion and a reduction of liver glycogen. Muscle glycogen and blood lactate levels were not altered by treatment with phenylephrine. Prazosin, 1 mg kg-1 s.c., partially attenuated phenylephrine-induced hyperglycaemia. Phenoxybenzamine infusion (16.6 micrograms kg-1 min-1) for 15 min suppressed the increase in blood glucose and the reduction in liver glycogen evoked by phenylephrine. This alpha-adrenoceptor blocker also clearly attenuated the blood glucose elevation observed on infusing adrenaline at 0.3 microgram kg-1 min-1. Blockade by phenoxybenzamine of phenylephrine- and adrenaline-induced hyperglycaemia was not accompanied by a significant increase in immunoreactive insulin plasma levels. Yohimbine infused at a rate of 20 micrograms kg-1 min-1, also completely blocked phenylephrine-induced hyperglycaemia. This suppressor effect was accompanied by a marked rebound in insulin secretion. It is concluded that in normal fasted rabbits stimulation of alpha-adrenoceptors induces hyperglycaemia. The increase in blood glucose depends mainly on liver glycogenolysis and inhibition of insulin secretion. Separate blockade of each component suffices to reduce alpha-adrenoceptor-mediated hyperglycaemia.

In vivo evidence for alpha 2-adrenergic-dependent inhibition of lipid mobilization in the rabbit.

Both alpha- and beta-agonists were infused i.v. in unanesthetized rabbits. The alpha-adrenergic effect on lipid mobilization was investigated by comparing the modifications of blood glycerol levels induced by clonidine (alpha 2-agonist) and amidephrine (alpha 1-agonist) alone or in combination with isoprenaline (beta-agonist). Clonidine (2 micrograms X kg-1 X min-1) was found to significantly reduce the basal or isoprenaline-stimulated glycerol levels whereas amidephrine (10 micrograms X kg-1 X min-1) was without any effect. Plasma IRI levels were also reduced by clonidine. The results suggest that the reduction of lipid mobilization produced by clonidine could have been the consequence of the stimulation of the alpha 2-adrenergic antilipolytic receptors previously shown to be present in isolated fat cell membranes.

Catecholamine-induced changes in plasma glucose and insulin levels in the unanesthetized rabbit.

Isoproterenol (ISO) produced dose-related increases in plasma immuno-reactive insulin (IRI) levels in unanesthetized rabbits, whereas epinephrine produced no significant changes. Dose rats of ISO or glucose that produced similar peak increases in glucose levels also produced similar increases in plasma IRI, but the responses to ISO occurred later and were of longer duration than the responses to glucose. Propranolol blocked the increase in plasma IRI produced by ISO, but the ISO-induced rise in plasma glucose was antagonized imcompletely. These data provide supporting evidence for the suggestion that the hyperglycemic activity of ISO in vivo is limited by concurrent ISO-induced changes in peripheral IRI levels.

The influence of propranolol on catecholamine-induced changes in carbohydrate metabolism in the rabbit.

In vitro evidence has indicated that isoproterenol (ISO) is more potent than epinephrine (EPI) in releasing glucose from rabbit liver slices and that propranolol (PROP) is a competitive antagonist of ISO. In contrast, dose-response data from fasted rabbits have shown that EPI is more potent than ISO in increasing plasma glucose levels and in lowering hepatic glycogen levels. Pretreatment with PROP abolished ISO-induced hyperglycemia and changes in liver and muscle glycogen levels whereas only the muscle glycogen depleting effect of EPI was altered significantly. These results suggest that factors other than stimulation of hepatic beta-receptors must be involved in EPI-stimulated depletion of liver glycogen and hyperglycemia in the intact rabbit.