ALBERT

Notes: 1 Pig nasal mucosal strips were incubated with α-adrenoceptor antagonists followed by α2-adrenoceptor agonist concentration–response curves. 2 Contractions elicited by the α2-adrenoceptor agonists BHT-920 (pD2 = 6.16 ± 0.07), UK 14,304 (pD2 = 6.89 ± 0.13) and PGE-6201204 (pD2 = 7.12 ± 0.21) were blocked by the α2-adrenoceptor antagonist yohimbine (0.1 μm). In contrast, the α1-adrenoceptor antagonist prazosin (0.03 μm) had no effect on the BHT-920-, UK 14,304- and PGE-6201204-induced contractions, but blocked the contractile response to the α1-adrenoceptor agonist phenylephrine (pD2 = 5.38 ± 0.04) and the mixed α1- and α2-adrenoceptor agonist oxymetazoline (pD2 = 6.30 ± 0.22). 3 The α2-adrenoceptor antagonist yohimbine (0.01–0.1 μm, pA2 = 8.04), α2B/C-adrenoceptor antagonist ARC 239 (10 μm, pKb = 6.33 ± 0.21), α2A/C-adrenoceptor antagonist WB 4101 (0.3 μm, pKb = 8.01 ± 0.24), α2A-adrenoceptor antagonists BRL44408 (0.1 μm, pKb = 6.82 ± 0.34) and RX 821002 (0.1 μm, pKb = 8.31 ± 0.35), α2C-adrenoceptor antagonists spiroxatrine (1 μm, pKb = 7.32 ± 0.32), rauwolscine (0.1 μm, pKb = 8.16 ± 0.14) and HV 723 (0.3 μm, pKb = 7.68 ± 0.14) inhibited BHT-920-induced contractions in pig nasal mucosa. 4 The present antagonist potencies showed correlations with binding affinity estimates (pKi) obtained for these antagonists at the human recombinant α2A- and α2C-adrenoceptors (r = 0.78 and 0.83, respectively) and with binding affinity estimates (pKd) obtained in pig native α2A- and α2C-monoreceptor assays (r = 0.85 and 0.78, respectively). No correlation was observed for the α2B-subtype. 5 In conclusion, contractile responses to phenylephrine, BHT-920, UK 14,304, PGE-6201204 and oxymetazoline indicate that α1- and α2-adrenoceptors are present and mediate vasoconstriction in pig nasal mucosa. Furthermore, correlation analysis comparing antagonist potency in pig nasal mucosa with affinities for human recombinant α2-adrenoceptors and native pig α2-adrenoceptors suggest that α2A- and α2C-adrenoceptor subtypes constrict pig nasal mucosa vasculature.

Notes: 1 The relationship between the effects of endogenous nitric oxide (NO) and prostanoids on the noradrenaline (NA)-induced contractions and the mechanisms involved were investigated in the rat perfused mesenteric bed, using NG-nitro-l-arginine methyl ester (l-NAME), a NO synthase inhibitor and sodium nitroprusside (SNP), a NO donor.2 The constrictor responses to NA were reduced to 50% by the cyclooxygenase inhibitor 10 μm indomethacin as well as by 1 μm SNP. When indomethacin and SNP were perfused simultaneously the contractions were further reduced.3 The NA-induced contractions were increased by the addition of 400 μml-NAME and the addition of either indomethacin or SNP abolished such increases. The simultaneous perfusion of both agents further reduced the contractions.4 Removal of the endothelium increased NA-induced contractions to a similar extent as l-NAME and this increase was abolished by indomethacin as well as by SNP.5 The perfusion of 10 μm NA augmented the release of prostaglandin (PG) F2α by the mesenteric bed without modifications in any other prostanoid. In the presence of l-NAME, this effect was further increased. However, SNP abolished the NA-induced stimulation of PGF2α release.6 In de-endothelialized preparations NA also stimulated PGF2α production as observed in intact preparations. This effect was more marked in the presence of l-NAME; in contrast, SNP abolished the stimulation.7 In conclusion, the present results suggest an opposite action between NO and PGF2α on the NA-induced contractions in the rat mesenteric bed.

Notes: 1 The present study was carried out to characterize the effect of adenosine on calcium dynamics in the rat portal vein. Isolated portal vein of male albino rats was used as the experimental model as it exhibits autorhythmicity. Adenosine and its analogues 2-CAD, N6-CHA and NECA were used to characterize the type of adenosine receptor involved and 2-CAD was used along with adenosine throughout the other part of study to characterize the effect of adenosine on Ca2+ dynamics.2 Adenosine and its analogues were found to inhibit the spontaneous contractions of rat portal vein in a concentration-related manner. The order of potency was NECA 〉 2-CAD 〉 N6-CHA 〉 adenosine. Incubation of the tissue with CGS∼15943A, an adenosine receptor antagonist, had a per se enhancing effect on autorhythmicity. Adenosine and 2-CAD failed to reverse the contractile response produced by hypertonic KCl (80 or 30 mm). Whereas adenosine and 2-CAD effectively relaxed the tissues contracted with phenylephrine (10−5 m).3 Preincubation of the tissue with 2-CAD (10−4, 10−5 or 10−6 m) for 5 min raised the threshold concentration of CaCl2 to evoke contractile response and also significantly increased the mean EC50 values of CaCl2. Nifedipine was found to be more potent than 2-CAD on Ca2+ channels.4 The results of the present study suggest that the endogenous adenosine plays a significant role in producing vascular relaxation through the participation of A2 receptor subtype. This effect may be due to its inhibitory effect on release of Ca2+ from the intracellular stores. Further to this effect, 2-CAD had a major inhibitory effect on voltage-operated Ca2+ channels compared with receptor-operated Ca2+ channels.5 It can be concluded that adenosine through its A2 receptor produces vasorelaxant effect by interfering with the release of Ca2+ from the intracellular stores coupled with influx of Ca2+ from the extracellular sources.

Notes: 1 Our previous report showed that in acute cholestasis, the subsensitivity to morphine inhibitory effect on electrical-stimulated contractions develops significantly faster in guinea-pig ileum (GPI) and in mouse vas deferens (MVD) (45.2 and 29.9 times, respectively) compared with non-cholestatic subjects. 2 The possible contribution of α2-adrenoceptor and nitric oxide (NO) pathways on the development of tolerance was assessed in GPI and MVD of cholestatic subjects. 3 Daily administration of naltrexone (20 mg kg−1), yohimbine (5 mg kg−1), and Nω-nitro-l-arginine methyl ester (l-NAME) (3 mg kg−1) to cholestatic animals significantly (P-value 〈 0.05) inhibited the process of subsensitivity in all groups. 4 Consistent with the literature, it was concluded that both the α2-adrenergic system and NO have close interaction with the opioid system and may underlie some of the mechanisms involved in the subsensitivity development to opioids in acute cholestatic states.

Notes: 1 We examined the role of the NO/cyclic GMP (cyclic GMP) pathway in nitric oxide (NO)- and vasoactive intestinal peptide (VIP)-induced relaxation of feline lower oesophageal sphincter (LES). Furthermore, it was studied whether methylene blue, LY83583 and ODQ, which are soluble guanylate cyclase (sGC) inhibitors, could inhibit NO-induced relaxation.2 The nitric oxide synthase (NOS) inhibitor, N-nitro-l-arginine (l-NNA) had no effect in sodium nitropruside (SNP)-induced relaxation, but 3-morpholinosydnonimine-N-ethylcarbamide (SIN-1)-induced relaxation was decreased by the pretreatment of l-NNA, which showed that SIN-1, not SNP, could activate NOS to cause relaxation. Methylene blue and LY83583 did not inhibit the relaxation by SNP and SIN-1. However, the more specific sGC inhibitor ODQ blocked the relaxation induced by NO donors.3 To identify the relationship of NOS, sGC and adenylate cyclase in VIP-induced relaxation, tissue were pretreated with l-NNA and ODQ and SQ22536. These inhibitors produced significant inhibition of this response to VIP. The adenylyl cyclase inhibitor SQ 22536 also inhibited relaxation by VIP.4 In conclusion, our data showed that SNP- and SIN-1-induced relaxation was mediated by sGC. Of sGC inhibitors, methylene blue and LY83583 were not adequate for the examination of NO donor-induced feline LES smooth muscle relaxation. VIP also caused relaxation by the pathway involving NO and cGMP and cAMP.

Notes: 1 The vasoconstrictor response to periarterial nerve electrical stimulation (PNS) and neurotransmission by ATP are discussed and illustrated, using canine isolated and perfused splenic arterial preparations. 2 The conditions for appearance of dominant purinergic constrictor response to PNS are discussed. 3 Modulation of the purinergic vasoconstrictor responses to PNS by several kinds of presynaptic receptor agonists and antagonists is reviewed. 4 Influences of purinergic responses to PNS by guanethidine, reserpine, tetrodotoxin (TTX) or ω-conotoxin GVIA (ωCTX) are also reviewed. 5 Effects of imipramine and removal of the endothelium are discussed. 6 Evidence is presented for selective inhibition of purinergic responses to PNS by an adequate cold storage of the vessel. 7 The roles of ATP released by PNS in isolated canine splenic arteries are proposed.

Notes: 1 The aim of the present study was to investigate the role of several possible neurotransmitters in mediating non-adrenergic, non-cholinergic (NANC) relaxation, and the effects of phosphodiesterase (PDE) III and V inhibitors on adrenergic and NANC relaxation in branch pulmonary artery (PA) of guinea-pig. 2 Under the NANC conditions, electrical field stimulation (EFS, 60 V, 0.2 ms, 20 Hz) induced a tetrodotoxin-sensitive relaxation of the histamine-precontracted PA rings. The nitric oxide (NO) synthase inhibitor NG-nitro-l-arginine methyl ester (l-NAME, 10−4 m) and the guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 10−5 m) partially inhibited the EFS-induced relaxation. The inhibitory effect of l-NAME was reversed completely by l-arginine (10−3 m), but not d-arginine (10−3 m). 3 This NANC relaxation was attenuated by 8-phenyltheophylline (10−5 m), a P1-purinoceptor antagonist. 4 The NANC response was potentiated by 10−6 m zaprinast, a type V PDE inhibitor, but was unaffected by 3 × 10−6 m milrinone, a type III PDE inhibitor. 5 Sodium nitroprusside (SNP) caused a concentration-dependent vasodilator effect which was potentiated by zaprinast, but unaffected by milrinone. Moreover, the effect of combination of zaprinast with milrinone was not significantly different from that observed with zaprinast alone. 6 Isoprenaline produced a concentration-dependent vasodilatation in branch PA of guinea-pig which was potentiated by both zaprinast and milrinone, the efficacy of milrinone being greater than zaprinast. 7 These results suggest that both nitrergic and purinergic pathways are involved in mediating the NANC relaxation in branch PA of guinea-pig. The combination of PDE III or V inhibitors with vasorelaxant drugs may be a hopeful approach for the treatment of pulmonary hypertension.

Notes: 1 One reason why rabbit jejunum is suitable for studying the mechanisms underlying the actions of the various neurotransmitters and their interactions is its spontaneous motility. The main regulator of spontaneous motility is the cholinergic system. How the cholinergic system regulates the spontaneous activity in the rabbit jejunum and how it affects the inhibitory action of α- and β-adrenoceptor agonists remains unclear. 2 We studied the influence of the cholinergic system and apamin-sensitive Ca2+-activated K+ channels on spontaneous contractions in the rabbit jejunum and on the inhibitory effects ofα1- and β-adrenoceptor agonists. 3 In naïve tissues, atropine (ATR, 7.4 × 10−8 m) and tetrodotoxin (8 × 10−8 m) almost completely inhibited – to a similar extent – the amplitude of spontaneous activity. Despite the presence of ATR or TDX, tissue contraction gradually recovered to about 50% of the baseline amplitude within 5–10 min. When ATR or TDX, respectively, were added to the TDX- or ATR-treated tissues, the recovered activity decreased weakly but significantly. After washout and a 45-min rest the contraction amplitude returned to baseline values. A further exposure to ATR or TDX reduced the contraction to a level significantly lower than the one obtained after TDX or ATR added 5 min after ATR or TDX, respectively. In preparations prestimulated for 10 min with acetylcholine (ACh), ATR abolished the TDX-resistant recovered spontaneous activity. 4 Adrenaline (ADR, 0.5–5 × 10−7 m) and phenylephrine (PHE, 1–10 × 10−7 m) inhibited tissue motility in naïve and in ATR- and in TDX-exposed preparations. But whereas in naïve preparations the α1-adrenoceptor antagonists completely antagonized inhibition induced by both drugs, in ATR- and TDX-exposed tissues they did so only partially for ADR. Agonist-induced inhibition had a rapid onset but rapidly faded; pendular movements took significantly longer to recover in ATR- and TDX-treated tissues than in naïve tissues. In tissues exposed for 2 min to ADR (0.5–5 × 10−7 m) or PHE (1–10 × 10−7 m), washout or addition of α1-adrenoceptor antagonists caused an immediate short-lasting increase in contraction amplitude. 5 Apamin (APAM, 5 × 10−9 m) caused a rapid and persistent increase in the amplitude of contractions. It also blocked the inhibitory responses to ADR and PHE, and removed washout-induced contractions. The APAM-induced increase in the contraction amplitude correlated with the increase obtained by washing out ADR or PHE. 6 Isoprenaline (at concentrations up to 2.8 × 10−7 m) produced no inhibitory response in naïve tissues, but it invariably blocked (at a concentration of 0.7 × 10−7 m) the recovered spontaneous activity (and sometimes depressed muscletone) in tissues exposed to ATR or TDX. Neither propranolol (3.4 × 10−7 m) nor APAM (5 × 10−9 m) counteracted these inhibitory effects. 7 These results indicate that spontaneous motility in the rabbit jejunum is predominantly mediated by neuronal release of ACh and by some other unidentified neuronal activity. Released ACh inhibits myogenic activity and strongly antagonizes β-adrenoceptor-induced APAM-insensitive inhibition but leaves α1 agonist-induced APAM-sensitive inhibition unchanged.

Notes: 1 This study used contractility methods with the portal veins of 5- and 14-week-old Wistar–Kyoto normotensive rats (WKY) and spontaneously hypertensive rats (SHRs). The SHRs are prehypertensive at 5 weeks. 2 The first part of our study was to determine whether the responsiveness to isoprenaline and forskolin was altered in the maturation of portal veins from normo- and prehypertensive rats. The responses to forskolin were similar on the portal veins of 5- and 14-week-old WKY and SHRs. 3 The sensitivity and maximum responses to isoprenaline were similar on portal veins of 5- and 14-week-old WKY. The sensitivity and maximum responses to isoprenaline were lower on the portal veins of 5-week-old SHRs (pD2 = 8.25, maximum = 85%) than age-matched WKY (pD2 = 8.79, maximum = 96%); these differences are not caused by hypertension. At 14 weeks, the sensitivity was similar (WKY pD2 = 8.74, SHR pD2 = 8.65) but the maximum responses to isoprenaline were lower on the portal veins SHRs (77%) than WKY (97%). Thus, the sensitivity to isoprenaline increases with the development of hypertension in the SHR portal vein. 4 The second part of the study was to determine whether the affinity for isoprenaline at β2-adrenoceptors and the fractional β2-adrenoceptor occupancy–response relationships on the portal vein were altered in maturation from normo- and pre-hypertensive rats. The effects of bromoacetylalprenololmenthane (BAAM), an irreversible β-adrenoceptor blocker, on the isoprenaline responses of 5- and 14-week-old WKY and SHRs were studied. Maturation of the WKY portal vein between 5 and 14 weeks was associated with a loss of affinity for isoprenaline (from pKA of 7.13 to 7.87), and increase in β2-adrenoceptor reserve (from 72 to 92% at the 95% response). There were similar affinity and reserve findings in the maturation of the SHR portal vein. Thus, there are major changes in β2-adrenoceptor structure and reserve in maturation on the portal vein that are irrespective of the development of hypertension.