A Dejonge

ORGANIC AND INORGANIC CALCIUM-ANTAGONISTS REDUCE VASOCONSTRICTION INVIVO MEDIATED BY POSTSYNAPTIC ALPHA-2-ADRENOCEPTORS

SummaryThe influence of various calcium antagonists and divalent metal cations on the pressor responses induced by the selective α1-adrenoceptor agonist methoxamine and the selective α2-adrenoceptor stimulating agent B-HT 920 (2-amino-6-allyl-5,6,7,8-tetrahydro-4H-thiazolo-[4,5-d]-azepine) was studied in pithed rats.1.The calcium antagonists verapamil, D 600 and nifedipine, when given intraarterially (i.a.) in doses up to 1 mg/kg did not influence the pressor effects of methoxamine. Only higher amounts of these calcium antagonistic drugs (1–3 mg/kg i.a.) somewhat reduced this pressor response.2.The vasoconstriction due to B-HT 920, as reflected by the increase in diastolic pressure, was markedly inhibited by verapamil, D 600 and nifedipine in a dose-dependent manner. In low doses a parallel displacement to the right was observed, whereas in higher amounts the shift was non-parallel.3.The divalent cations Mn2+, Ni2+ and Co2+ (0.05–0.15 mmol/kg i.a.) hardly affected the pressor effect of methoxamine, whereas B-HT 920-induced vasoconstriction was highly sensitive to these metal ions. La3+ and Mg2+ were ineffective.4.The calcium antagonists verapamil, D 600 and nifedipine displayed only minor affinities for [3H]prazosin (α1) as well as [3H]clonidine (α2) binding sites of rat brain membranes.5.It is concluded that an influx of extracellular Ca2+ is necessary for the vasoconstriction in vivo initiated by stimulation of vascular postsynaptic α2-adrenoceptors. On the other hand, vasopressor responses to α1-adrenoceptor stimulation are not directly dependent on a transmembrane influx of calcium ions.

DIFFERENTIAL EFFECT OF CALCIUM ENTRY BLOCKERS ON ALPHA-1-ADRENOCEPTOR-MEDIATED VASOCONSTRICTION INVIVO

SummaryThe effects of the calcium entry blockers nifedipine, (−)-verapamil and the dihydropyridine derivative PY 108-068 were evaluated on the increase in diastolic pressure of pithed normotensive rats caused by the selective α1-adrenoceptor agonists cirazoline, (−)-phenylephrine, (±)-erythro-methoxamine, (−)-amidephrine and St 587 [(2-chloro-5-trifluoromethylphenylimino)-2-imidazolidine] as well as by the mixed α1/α2-adrenoceptor agonists clonidine and DPI [(3,4-dihydroxyphenylimino)-2-imidazolidine]. The calcium entry inhibitors (up to 3 mg/kg) caused 3- to 5-fold, parallel rightward shifts of the log dose-pressor effect curves to cirazoline, (−)-phenylephrine, (±)-erythro-methoxamine and (−)-amidephrine accompanied by only a slight depression of the maximal pressor response. In contrast, the calcium entry inhibitors produced a dose-dependent profound depression of both maximum and slope of the log dose-pressor response curves to St 587 and clonidine. For DPI about 10-and 100-fold parallel displacements to the right without reduction of the maximum were found following treatment with 1 and 3 mg/kg of nifedipine, respectively. Infusion of vasopressin to counteract the vasodilatory action produced by the calcium entry inhibitors did not significantly change the pattern of interference observed under the conditions of decreased baseline diastolic pressure. The results indicate that α1-adrenoceptor-mediated vasoconstriction in the pithed normotensive rat, which is characterized by its sensitivity to blockade by prazosin and its relative insensitivity to antagonism by yohimbine or rauwolscine, can be subdivided into two distinct processes which are differentially influenced by blockade of calcium entry. It is suggested that cirazoline, (−)-phenylephrine, (±)-erythro-methoxamine and (−)-amidephrine initiate vasoconstriction predominantly by a mechanism independent of extracellular calcium influx. On the other hand, St 587 and clonidine increase arterial pressure via a mechanism which seems to be mainly governed by an entry of extracellular calcium ions. DPI probably triggers both processes of vasoconstriction in the intact circulatory system of the pithed normotensive rat.

Captopril impairs the vascular smooth muscle contraction mediated by postsynaptic α2-adrenoceptors in the pithed rat

Since two different types of α-adrenoceptors (α1- and α2-) subserve vasoconstriction in pithed rats and noradrenaline behaves as a non-selective agonist of both receptors (for review see Timmerman and van Zwieten, 1981), we have investigated this interaction between noradrenaline and captopril in more detail to identify the class of α-adrenoceptor involved. This study confirms the findings reported recently (Antonaccio and Kerwin, 1981; Clough et al., 1981) that captopril inhibits postjunctional vasocontrictor responses to noradrenaline in pithed rats. However, captopril interfered with the vasoconstriction elicited by noradrenaline only in those animals in which the vascular postsynaptic α2-adrenoceptor system could be triggered (saline-and prazosin-treated rats). Captopril was without effect after removal of the contribution of post-synaptic α2-adrenoceptor stimulation to the in vivo vascular smooth muscle contraction due to (-)-noradrenaline (rauwolscine-treated rats). These results indicate that captopril selectively impairs the vasoconstriction subserved in vivo by postjunctinal α2-adrenoceptors. The vasoconstrictor process initiated by α1-adrenoceptor activation is not affected by captopril.

EFFECT OF CAPTOPRIL ON SYMPATHETIC NEUROTRANSMISSION IN PITHED NORMOTENSIVE RATS

Captopril significantly diminished the basal diastolic blood pressure and the vasopressor response to electrical stimulation of the thoracic-lumbar spinal cord in pithed normotensive rats. The reduction of the hypertensive response to electrical stimulation was more pronounced after bilateral adrenalectomy. Captopril also diminished the vasopressor response to intravenously administered (-)-noradrenaline. Pretreatment of the animals with indomethacin had no effect on the vasopressor response to electrical stimulation and did not affect the sympathoinhibition of captopril. After bilateral nephrectomy, 18-24 h previously, the basal diastolic blood pressure and the vasopressor response to electrical stimulation were reduced and captopril had no additional inhibitory effect on these parameters. In indomethacin-pretreated animals with intact kidneys, restoration of the basal diastolic blood pressure with angiotensin II (AII) completely abolished the sympathodepressive effect of captopril. When the reduction in basal diastolic blood pressure with captopril was prevented by vasopressin, converting enzyme inhibition had no depressive effect on the hypertensive response to intravenously administered (-)-noradrenaline and did not influence sympathetic neurotransmission in animals with intact adrenals. However, a small, but significant reduction of the hypertensive response to electrical stimulation by captopril was still detectable in bilaterally adrenalectomized rats. The results suggest that endogenous AII facilitates sympathetic neurotransmission in vascular smooth muscle of pithed normotensive rats. However, the modulatory action of endogenous AII largely results from an effect on basal arteriolar smooth muscle tone and should, therefore, be considered as non-specific facilitation. Genuine prejunctional facilitation of the sympathetic neurotransmission in vascular smooth muscle can only be observed after bilateral adrenalectomy but this effect of endogenous AII appears of minor significance, at least in pithed normotensive rats.

Invariable susceptibility to blockade by nifedipine of vasoconstriction to various alpha 2-adrenoceptor agonists in pithed rats.

The sensitivity of the increase in diastolic pressure brought about by the selective agonists of α2‐adrenoceptors, B‐HT 920, B‐HT 933, xylazine, UK‐14,304, M‐7, TL‐99 and DP‐6, 7‐ADTN in pithed normotensive rats to blockade by the calcium entry inhibitor nifedipine has been investigated. To exclude any participation of vascular α1‐ and β2‐adrenoceptors, as well as cardiac β1‐adrenoceptors, in the pressor responses, the study was made after treatment of the pithed rats with prazosin (0·1 mg kg−1) and (‐)‐propranol (1 mg kg−1). Without exception, the preferential agonists of α2‐adrenoceptors elicited vasoconstrictor responses which were susceptible to inhibition by nifedipine (0·03–1 mg kg−1) in a dose‐dependent manner regardless of the differences in intrinsic activity of the compounds. The pressor activity was almost completely abolished after 1 mg kg−1 of nifedipine. The results show that vasoconstriction induced in pithed rats by various selective stimulating agents of postjunctional vascular α2‐adrenoceptors is invariably and equally sensitive to attenuation by nifedipine. This susceptibility of α2‐adrenoceptor‐mediated vasoconstriction to impairment by blockade of calcium entry is not dependent on the nature, the potency or the efficacy of the agonist.

INTERACTIONS BETWEEN THE PUTATIVE CALCIUM ENTRY PROMOTOR BAY-K-8644 AND PRESSOR-RESPONSES PRODUCED BY ALPHA-1-ADRENOCEPTOR AND ALPHA-2-ADRENOCEPTOR AGONISTS IN THE PITHED NORMOTENSIVE RAT

SummaryInteractions between the putative calcium entry promotor Bay k 8644 and both α-1 and α1-adrenocepter mediated increases in diastolic pressure were studied in the pithed normotensive rat. The α1-adrenocepter mediated pressor responses elicited by B-HT920, TL-99, DP-6,7-ADTN and B-HT958 were potentiated by Bay k 8644, reflected by a leftward shift and an increase in the maximum of the log dose-pressor respinse curves. The α-1-adrenocepter effects elicited by cirazoline, methoxamine, (−)-amidephrine, St 587, (−)-phenylephrine and Sgd 101/75 were less enhanced by Bay k 8644. Only a leftward shift of the dose-response curves was observed, which was most pronounced for (−)-phenylephrine and Sgd 101/75. The α-1 and α2-adrenocepter pressor components of (−)-noradrenaline were similarly distinguished by Bay k 8644 as observed for the selective α-1 or α2-adrenocepter agonists.Effects of Bay k 8644 on the increase in diastolic pressure mediated by B-HT 920, St 587 and cirazoline were also studied after pretreatment with the calcium entry blocker nifedipine. After additional pretreatment with nifedipine the potentiation by Bay k 8644 observed for B-HT 920 and St 587 was more pronounced. The presence of nifedipine had no effect on the interaction between Bay k 8644 and cirazoline.It is concluded that Bay k 8644 behaves as a mirror image of nifedipine. The observation that Bay k 8644 enhances α2-adrenocepter mediated pressor effects more effectively than α1-adrenocepter increases in diastolic pressure is in accordance with the hypothesis of the more pronounced calcium dependency of α2-adrenocepter mediated pressor responses. The data obtained for ceptor mediated pressor responses. The data obtained for St 587 and (−)-phenylephrine are in apparent contradiction to the finding that the pressor responses to the former drug are more markedly inhibited by calcium entry blockade than those of the latter. It is suggested that St 587 employs calcium channels which are already maximally modulated and that (−)-phenylephrine makes use of calcium channels which are in a rather inactive state. The hypothesis is put forward that the intrinsic activity of α2-adrenocepter agonists reflects their ability to bring calcium channels in an active state.

α1-adrenoceptor-mediated vasoconstriction in vivo to enantiomers of SK&F 89748-A

The pressor activity of the 1-enantiomer of SK & F 89748-A, 1,2,3,4-tetrahydro-8-methoxy-5-(methylthio)-2-naphthalenamine, in pithed normotensive rats was found comparable with that of 1-phenylephrine. The d-enantiomer was half as potent. The log dose-pressor effect curves for d- and 1-SK & F 89748-A were not influenced by reserpine treatment (2 X 5 mg/kg i.p., -48 and -24 h), were virtually unaffected by yohimbine (1 mg/kg i.v., -15 min) but were markedly shifted to the right by prazosin (0.1 mg/kg i.v., -15 min) and phentolamine (1 mg/kg i.v., -15 min). Similar observations were made for the 1-enantiomer in pithed cats. It is concluded that d- and 1-SK & F 89748-A are potent, directly acting highly selective agonists of (vascular) postjunctional alpha 1-adrenoceptors. Potency and selectivity were equally pronounced for both enantiomers. The currently available selective agonists of alpha 1-adrenoceptors, including the optical isomers of SK & F 89748-A, cannot distinguish between alpha 1- and alpha 2-adrenoceptors. This conclusion is based on binding affinity since these affinities are linearly correlated as shown by radioligand displacement experiments.

Role of ganglionic M-1 and M-2 receptors in the neuronal control of the cardiovascular system of the normotensive rat as determined with pilocarpine

The adrenoceptors involved in the increase in diastolic pressure and heart rate elicited by i.v. administration of pilocarpine to the pithed rat were assessed using as pharmacological tools the alpha 1-adrenoceptor antagonist prazosin, the alpha 2-adrenoceptor antagonist rauwolscine, the beta 1-adrenoceptor blocker atenolol and the beta 2-adrenoceptor blocker ICI 118,551. Pilocarpine indirectly activated vascular alpha 1- and cardiac beta 1-adrenoceptors. By using the M-1 antagonist pirenzepine and the mixed M-1/M-2 antagonist dexetimide, pilocarpine was shown to be a mixed M-1/M-2 agonist. Pilocarpine initiated antagonistic effects on intrasynaptic alpha 2-adrenoceptor-mediated pressor responses and not on those triggered by extrasynaptic alpha 2-adrenoceptors. During vasopressin infusion to counteract a possible vasodilator action of pilocarpine, it was demonstrated that pilocarpine indirectly activated alpha 1- and alpha 2-adrenoceptors. The results support the hypothesis that intra- and extrasynaptic alpha 2-adrenoceptors comprise different populations and that the neuronal control of alpha 2-adrenoceptors is mediated by ganglionic M-2 receptors.

EFFECT OF ADRENALECTOMY AND DEMEDULLATION OF THE ADRENALS ON VASOPRESSOR RESPONSES TO STIMULATION OF POSTJUNCTIONAL ALPHA-2-ADRENOCEPTORS

After adrenalectomy performed 18-12 h previously, the vasopressor responses induced by stimulation of postjunctional alpha 2-adrenoceptors (agonist: B-HT 920) were attenuated in pithed normotensive rats in contrast to pressor responses mediated by postjunctional alpha 1-adrenoceptors (agonist: cirazoline). Aldosterone and desoxycorticosterone both enhanced the vasopressor responses to the alpha 2-adrenoceptor agonist B-HT 920 in adrenalectomized rats, but did not fully restore the effects. Demedullation of the adrenals also attenuated the vasoconstrictor processes evoked by stimulation of postjunctional alpha 2-adrenoceptors 18-22 h later. This effect was reversed after an infusion with adrenaline. The combination of aldosterone and adrenaline produced full recovery of the increase in diastolic pressure elicited via alpha 2-adrenoceptors in adrenalectomized (18-22 h) animals. The results suggest that circulating adrenaline and aldosterone (or corticosterone) are required for optimal functioning of vascular postjunctional alpha 2-adrenoceptors.

LACK OF RELATIONSHIP BETWEEN INTRINSIC ACTIVITY AND SUSCEPTIBILITY OF PRESSOR-RESPONSES TO BLOCKADE BY NIFEDIPINE AMONG THE ALPHA-2-ADRENOCEPTOR AGONISTS B-HT-920 AND B-HT-958

SummaryFollowing i.v. bolus injections into pithed normotensive rats, the maximal diastolic pressor responses to B-HT 920 and B-HT 958 amounted to 115 and 35 mm Hg, respectively. Prazosin (0.1 mg/kg, i.v.,-15 min) was without effect on the log dose-pressor effect curve of B-HT 958, whereas yohimbine (1 mg/kg, i.v.,-15 min) shifted this curve about 30-fold to the right, showing the exclusive participation of α2-adrenoceptors in the vasoconstrictor response to B-HT 958. In doses of 10 and 30 mg/kg, B-HT 958 displaced the log dose-vasoconstrictor effect curve of B-HT 920 approximately 6- and 30-fold, respectively, to the right, illustrating the partial agonism of B-HT 958 at postjunctional vascular α2-adrenoceptors. Despite the marked difference in intrinsic activity of B-HT 920 and B-HT 958, the calcium entry blocker nifedipine exhibited a comparable inhibitory action on the vasopressor responses to both agonists. This finding indicates that partial and full agonism at vascular α2-adrenoceptors are not related to the susceptibility of the initiated pressor response to inhibition by calcium entry blockade.