Contraction Of Human Coronary Arteries Research Articles

Pharmacological characterization of tegaserod at the wild type and 124Cys variant of the human 5-HT1B receptor

Sumatriptan, an antimigraine drug, causes contraction of human coronary arteries through activation of 5-HT1B receptors which couple to Gi/Go inducing inhibition of adenylate cyclase. At a rare, naturally occurring human receptor variant (124Cys-h5-HT1B), sumatriptan has previously been shown to act as a more potent agonist than at wild-type receptor. Tegaserod, a 5-HT4-receptor agonist, developed for the treatment of functional gastrointestinal disorders, has been suspected to be involved in very rare cardiac ischemic events in patients with cardiovascular risk factors. In this study, we examined the potential agonist-like effects of tegaserod in comparison with sumatriptan at heterologously expressed human wild type and 124Cys-variant 5-HT1B receptors, using assays addressing G-protein coupling and inhibition of forskolin-stimulated cyclic AMP accumulation. Sumatriptan exhibited agonist effects as previously reported, whereas tegaserod acted as partial agonist at both wild type and 124Cys-variant h5-HT1B receptors (expressed in rat C6 glioma cells). Sumatriptan and tegaserod were more potent at the 124Cys-variant h5-HT1B receptor. It remains to be shown whether the very rare cardiovascular side effects reported with these drugs are predominantly observed in patients homozygously expressing the variant receptor.

Effects of SL 65.0472, a novel 5-HT receptor antagonist, on 5-HT receptor mediated vascular contraction

5-Hydroxytryptamine (5-HT) contracts vascular smooth muscle and pharmacological and molecular biological data suggest that these effects are mediated primarily by stimulation of 5-HT 1B and 5-HT 2A receptor subtypes. We have studied the properties of 7-fluoro-2-oxo-4-[2-[4-(thieno[3,2- c] pyridin-4-yl) piperazin-1-yl] ethyl]-1,2-dihydroquinoline-1-acetamide (SL 65.0472 ), a novel 5-HT receptor antagonist, in isolated vascular preparations contracted by 5-HT or sumatriptan. In canine isolated saphenous vein strips (putatively 5-HT 1B-mediated contraction), SL 65.0472 antagonised sumatriptan-induced contractions in a competitive manner (p A 2 8.17±0.36). 5-HT contracts rabbit aorta by stimulation of 5-HT 2A receptors. SL 65.0472 displaced the 5-HT concentration response curve in rabbit aorta rightwards with a significant reduction in maximum. The apparent p K B value was 8.58±0.18. 5-HT-induced contractions of human coronary arteries are mediated by a mixed population of 5-HT 1B and 5-HT 2A receptors. SL 65.0472 produced rightward parallel shifts of the 5-HT concentration response curves in all tissues studied (p A 2 8.8±0.14, n=7). In conclusion, SL 65.0472 is a potent antagonist of vascular smooth muscle contraction in vitro mediated by 5-HT receptor stimulation.

Characterisation of 5-HT receptors in human coronary arteries by molecular and pharmacological techniques

5-Hydroxytryptamine (5-HT) can produce both vasoconstrictor and vasorelaxant effects in human coronary arteries and the response to 5-HT can be influenced by the presence of disease. The aim of the present study was to elucidate the 5-HT receptor subtypes responsible for mediating 5-HT-evoked contraction of human coronary arteries using pharmacological, molecular and immunocytochemical approaches. Normal human coronary arteries, with intact endothelium, were mounted in tissue baths, and the vascular responses to 5-HT and 5-HT receptor agonists were studied. The effects of 5-HT 1 and 5-HT 2 receptor antagonists on these responses were also studied. Expression of messenger ribonucleic acid (mRNA) encoding different 5-HT receptors in human coronary arteries, atrium, ventricle wall and epicardium was determined using reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot analysis. The expression of 5-HT 1B or 5-HT 1D receptor protein was studied using subtype selective antibodies and standard immunocytochemical techniques. The rank order of 5-HT receptor agonist potency in causing vasoconstriction was 5-carboxamido tryptamine, (5-CT)>zolmitriptan=BW183C91 ( N 10-desmethyl zolmitriptan)=α-methyl-5-hydroxytryptamine (α-CH 3-5-HT)=5-HT=sumatriptan>2-methyl-5-hydroxytryptamine (2-CH 3-5-HT)=8-hydroxy-DPAT (8-OH-DPAT). α-CH 3-5-HT, 5-CT, 5-HT, zolmitriptan and BW 183C91 were significantly more potent (approximately 3-fold) than sumatriptan and 2-CH 3-5-HT, which in turn were more potent than 8-OH-DPAT. Ketanserin and methiothepin (5-HT 2 and 5-HT 1 receptor antagonists, respectively) caused parallel rightward shifts of the concentration–effect curves to α-CH 3-5-HT or 5-CT, respectively, without changing the maximum contractile response. In human coronary arteries, atrium, ventricle and epicardium. RT-PCR products corresponding to the human 5-HT 2A, 5-HT 1B and 5-HT 1F receptors were expressed in high levels, mRNAs coding for 5-HT 7, 5-HT 1A and 5-HT 1D receptors were only weakly expressed. No 5-HT 1E receptor mRNA was detected. In coronary arteries there was a differential expression of 5-HT 1B versus 5-HT 1D receptor mRNAs, with 5-HT 1B mRNAs being found in greater abundance. Dense 5-HT 1B-immunoreactivity was detected on smooth muscle layer within coronary artery, however, 5-HT 1D-immunoreactivity was not detected. It is concluded that 5-HT-evoked contraction of human coronary arteries is most probably mediated via the activation of both 5-HT 1B and 5-HT 2A receptors.

Up-Regulation of Endothelin-B Receptors in Atherosclerotic Human Coronary Arteries

Both endothelin-A (ETA) and endothelin-B (ETB) receptors are known to be present in human coronary arteries. However, their absolute and relative amounts, functional roles, and the influence of pathology are uncertain. The goal of the present study was to characterize endothelin receptors mediating constriction in human coronary arteries and to assess the influence of cardiomyopathy (CMP) and coronary artery disease (CAD) on ET receptors in human tissue. For comparison, porcine coronary arteries were evaluated in parallel. Competition binding experiments using [125I]ET-1 and different selective and nonselective ETA- and ETB-receptor agonists or antagonists revealed similar relative densities (relative Bmax) of ETA and ETB receptors in coronary arteries from human cardiomyopathic hearts (83% ETA and 17% ETB; n = 5) and porcine hearts (78% ETA and 22% ETB; n = 5). In marked contrast, the relative Bmax of ETB receptors were significantly higher in coronary arteries from human atherosclerotic hearts (51% ETA and 49% ETB; n = 3). Total receptor density (Bmax; fmol/mg protein) was highest in porcine (385 +/- 29) arteries, followed by human CAD (253 +/- 41) and CMP (174 +/- 20) coronary arteries. The relative and absolute Bmax values for ETA and ETB receptors in coronary arteries from a donor heart were similar to those obtained in CMP hearts. There were no significant differences in affinity constants (KD) values for ET-1, ET-3, Sarafotoxin S6c (SRTX S6c), BQ-123, and bosentan (Ro 47-0203) between tissues. In human coronary arteries from CMP hearts, ET-induced constriction seemed to be solely mediated via ETA receptors. In contrast, in porcine coronary arteries 20% of the maximal effect mediated by ET-1 could be attributed to ETB receptors, in agreement with the binding data. The functional role of ETB receptors in CAD tissue could not be evaluated because of the occurrence of spontaneous phasic contractions. We conclude that ETB receptors are up-regulated in human atherosclerotic coronary arteries. Further studies are needed to determine the pathophysiological importance of these receptors.

Role of Endothelin in Spontaneous Phasic Contraction of Human Coronary Arteries

Isolated human coronary arteries are known to develop spontaneous phasic contractions (SPCs). The goal of the present study was to assess the role of endothelin (ET) in these contractions. Endothelium-denuded rings from left anterior descending (LAD) coronary arteries of patients with coronary artery disease (CAD; n = 3) undergoing cardiac transplantation developed SPCs within 1-2 h. The contractions were abolished by the ETA receptor antagonist BQ-123 or the mixed ET receptor antagonist bosentan (1 and 3 microM), whereas the contractions persisted for several hours in control rings. Exogenous ET-1 (10 pM) increased the amplitude of SPCs and initiated SPCs in rings that did not exhibit SPCs. Higher concentrations of ET-1 (> or = 1 nM) were able to re-induce SPC in rings in which SPCs had been abolished by BQ-123 or bosentan. These results suggest that ET plays an important role in spontaneous phasic contractions of isolated human coronary artery rings.

Role of Endothelin in Spontaneous Phasic Contraction of Human Coronary Arteries

Role of Endothelin in Spontaneous Phasic Contraction of Human Coronary Arteries

Postmortem functional changes in coronary and cerebral arteries from humans and monkeys.

Contractile responses to 30 mmol . litre-1 K+ of helical strips of coronary arteries from human cadavers did not change within 5 h after death; however, they were suppressed 8 h after death. In coronary arteries from monkey cadavers, the K+-induced contractions did not significantly differ within the first 5 h, but were suppressed 12 h after death. On the other hand, K+-induced contractions were retained without deterioration in cerebral artery strips from human cadavers 20 to 24 h after death and those from monkey cadavers 8 to 16 h. Acetylcholine caused contractions of human coronary arteries, but caused only a relaxation of monkey coronaries which was abolished by rubbing off the endothelium. These responses were attenuated by no more than K+-induced contractions up to 12 h after death. Maximum contractions induced by noradrenaline, histamine and serotonin remained the same in human coronary arteries for 3 to 5 h after death. Similar magnitudes of contraction were elicited by noradrenaline in human cerebral arteries up to 20 h after death. It appears that the reactivity of human coronary arteries to K+ and other vasoconstrictor agents used is normally retained for at least 6 h after death and that of human cerebral arteries up to 24 h.

Cholinergic mechanisms in human coronary artery preparations: implications of species differences.

Acetylcholine dilates most arteries, including dog coronaries, if the endothelium is intact. The present study has shown only contraction of human coronary arteries to acetylcholine. Both strip and ring preparations of human coronary epicardial vessels, the latter done particularly to protect the intimal surface from unintentional denudation, contracted to acetylcholine at low to high concentrations (6.84 X 10(-9)-2.05 X 10(-5) M). These responses were blocked by atropine (3.45 X 10(-6) M). Acetylcholine contracted the arteries about as much as ergonovine and considerably more than noradrenaline. Field stimulation of coronary artery strips caused a vasoconstriction which was partially antagonized by atropine (3.45 X 10(-6) M). The release of [3H]noradrenaline from superfused coronary artery preparations during field stimulation was inhibited by methacholine (6.24 X 10(-6) M), a stable muscarinic analogue of acetylcholine. Dog coronary arteries relaxed to acetylcholine but not if the endothelium was intentionally denuded, in which case there was either no response at all or a weak relaxation. Coronary arteries of sheep, pig and cattle always contracted to acetylcholine, and those of monkey contracted in two out of three responsive preparations. Histological examination of the intimal surface of human coronary vascular segments confirmed the presence of an intact endothelial cell layer. Rabbit aorta gave dilator responses to acetylcholine even after being left in the animal for as long after death as the human arteries had been; they did not give dilator responses after the endothelium was rubbed off. It is concluded that cholinergic vasoconstriction of coronary arteries occurs in humans, though not in the dog, and is probably important in some cases of coronary artery spasm.

Isolated human coronary arteries in response to vasoconstrictor substances.

In helical strips of human epicardial coronary arteries, norepinephrine produced a concentration-related contraction; the contractions relative to those induced by 30 mM K+ were greater in the proximal portion of the arteries than in the distal portion. The amine-induced contraction was suppressed by treatment with phentolamine. Acetylcholine contracted human coronary arteries but, in contrast, relaxed the monkey coronary arteries (both freshly excised and cadaver) previously contracted with prostaglandin F2 alpha. Both the contraction and relaxation induced by acetylcholine were suppressed by atropine. Removal of the endothelium abolished the relaxation of monkey arteries but did not significantly alter the contraction of human arteries. Human coronary arteries responded to histamine with contractions, which were reversed to relaxations following treatment with chlorpheniramine. It is concluded that, as far as the portions of human coronary arteries used in the present study are concerned, the arterial contraction mediated via alpha-adrenoceptors is inversely related to the distance from the coronary artery orifice. Acetylcholine produces contractions of human coronary arteries, possibly due to activation of muscarinic receptors on smooth muscle cells. Histamine-induced contractions appear to be mediated via H1-receptors.