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T M Griffith

Publications and source records attributed to T M Griffith.

88 records · Page 5Linked to original sources

Influence of endothelium on drug-induced relaxation of the rabbit aorta.

The influence of endothelium on relaxation induced by glyceryl trinitrate (GTN) and isoprenaline (I) has been studied in isolated rabbit aortic ring preparations, in the resting state and after constriction with 5-hydroxytryptamine (5HT). In resting preparations relaxant responses to GTN and I were smaller in the presence of endothelium. In preparations constricted with 5HT relaxant responses to GTN and I were conversely greater in the presence of endothelium. These paradoxical findings are discussed in relation to basal release of endothelium derived relaxant factor.

Animals↗

Production of endothelium derived relaxant factor is dependent on oxidative phosphorylation and extracellular calcium.

The production (synthesis or release or both) of endothelium derived relaxant factor was studied in rabbit aortic strip preparations and an aortic-coronary artery bioassay system. Production of endothelium derived relaxant factor was rapidly inhibited by agents that inhibit mitochondrial electron transport or F1-ATPase, or which uncouple oxidative phosphorylation, but was only slowly impaired by inhibition of glycolysis. It was dependent also on the presence of extracellular calcium with a rapid on-off response time. This study shows that production of endothelium derived relaxant factor appears to be dependent on both oxidative phosphorylation and extracellular calcium.

Acetylcholine↗

Haptoglobin-haemoglobin complex in human plasma inhibits endothelium dependent relaxation: evidence that endothelium derived relaxing factor acts as a local autocoid.

Endothelium dependent relaxation of isometrically mounted rabbit aortic strip preparations was rapidly inhibited by human plasma at dilutions down to 1:1000. Gel filtration and ion exchange chromatography were used to demonstrate that this inhibitory activity was present in fractions containing haptoglobin. Purified haptoglobin itself possessed no inhibitory action against endothelium dependent relaxation, but the haptoglobin-haemoglobin complex did, consistent with the documented ability of haemoglobin to inhibit this phenomenon. The concentration of haemoglobin normally bound to haptoglobin is sufficient to account for the inhibitory properties of human plasma. This suggests that endothelium derived relaxing factor exerts no downstream intravascular effect in vivo and thus that its physiological dilator role is that of a local autocoid acting on subjacent smooth muscle.

Animals↗

Endothelium-derived relaxing factor alters calcium fluxes in rabbit aorta: a cyclic guanosine monophosphate-mediated effect.

1. Measurement of tension and 45Ca influx and efflux were used to study the effects of endothelium-derived relaxing factor (EDRF), sodium nitroprusside and 8-bromo-cyclic guanosine monophosphate (GMP) on contractile responses and calcium movements in aortic ring preparations of the rabbit. 2. EDRF activity, induced by stimulating endothelium-containing rings with acetylcholine, was associated with relaxation of noradrenaline-constricted rings and with a marked reduction of noradrenaline-stimulated increase in calcium influx. Sodium nitroprusside and 8-bromo-cyclic GMP had a similar effect in de-endothelialized preparations. 3. EDRF also inhibited noradrenaline-stimulated calcium efflux. Sodium nitroprusside and 8-bromo-cyclic GMP had a similar effect in de-endothelialized preparations, both in the presence and absence of extracellular calcium. 4. The vascular smooth muscle relaxant effect of EDRF and of nitrovasodilators may be effected by a cyclic GMP-mediated reduction of cytosolic calcium, through both inhibition of calcium influx and reduction of intracellular calcium release.

8-Bromo Cyclic Adenosine Monophosphate↗

Evidence that cyclic guanosine monophosphate (cGMP) mediates endothelium-dependent relaxation.

The mechanism of action of endothelium-derived relaxant factor (EDRF) was studied using aortic strip preparations of the rabbit and a bioassay system of a rabbit coronary artery perfused in series with an intact aorta. Methylene blue (an inhibitor of guanylate cyclase) inhibited, and 2-O-propoxyphenyl-8-azapurine-6-one (MB22948, an inhibitor of cGMP phosphodiesterase) potentiated the vascular effects of EDRF whether these were due to its basal or to stimulated release. Infusion of these agents at different sites in the bioassay indicated that they act pharmacologically at the smooth muscle level and not on release of EDRF or by chemical interaction with EDRF. The data are consistent with the hypothesis that EDRF-induced relaxation is mediated by elevation of smooth muscle cGMP levels.

Animals↗

Studies of endothelium-derived relaxant factor (EDRF), its nature and mode of action.

The effect of endothelium on constrictor responses to 5-hydroxytryptamine, histamine, phenylephrine and acetylcholine was studied and shown to be much greater in isolated perfused coronary arteries than aortic strips of the rabbit. Localised endothelial damage predisposed nonspecifically to 'coronary spasm'. Endothelium-dependent dilatation was shown by bioassay to be mediated by a humoral agent, endothelium-derived relaxant factor (EDRF), with half-life of 6 s. Experiments with inactivating agents indicate that EDRF is not a cyclo-oxygenase or lipoxygenase product and not a free radical; they imply that it contains a carbonyl group at or near its active site. Experiments in which guanylate cyclase and cGMP phosphodiesterase were inhibited indicate that EDRF acts by elevating smooth muscle cGMP. Ergometrine was shown to stimulate EDRF activity which may be relevant to its clinical use in provoking coronary spasm. The physiological role and pathophysiological relevance of this novel, ubiquitous and potent endogenous vasodilator are not yet known; it may be of particular importance in modulating coronary vasomotor responses.

Acetylcholine↗

Endothelium influences coronary and aortic vasomotion by release of an unstable humoral factor.

Using an isolated perfused coronary-artery preparation, we have demonstrated the ability of endothelium inhibit markedly to vasomotion in rabbit coronary arteries. Using a bioassay system, we have shown this effect to be mediated via the release of an unstable humoral agent (t 1/2 approximately equal to 6 sec) from endothelial cells, and we have partially characterized its chemical nature.

Acetylcholine↗

Ergometrine-induced arterial dilatation: an endothelium-mediated effect.

Ergometrine has generally been regarded as a vasoconstrictor and is used clinically to provoke coronary vasospasm in susceptible patients [3, 8, 9]. The ergometrine response appears however to be complex in that it can be biphasic, the constrictor response being preceded by an initial dilator response in experimental models [4]. The explanation for this is unknown, as is the underlying mechanism responsible for the clinical condition of coronary vasospasm. We have investigated this biphasic response in isolated artery preparations and shown that the dilator component is due to ergometrine-induced release of endothelium-derived relaxant factor (EDRF).

Animals↗

Isolated perfused rabbit coronary artery and aortic strip preparations: the role of endothelium-derived relaxant factor.

Isolated perfused coronary arteries and aortic ring preparations of rabbits were studied, both with intact endothelium and with endothelium removed by K-rich solution and friction respectively. Constrictor dose-responses to histamine, acetylcholine, phenylephrine and 5-hydroxytryptamine (5-HT) were measured. They were greatly depressed by the presence of endothelium in coronary preparations. In aortic preparations endothelium affected dose-responses relatively little, depressing the response to acetylcholine but apparently increasing the responses to the other three agents. Acetylcholine relaxed pre-constricted coronary or aortic preparations but only when endothelium was present. This relaxation was inhibited by quinacrine or hydroquinone. Aortic preparations had resting tone which could be increased by hydroquinone if endothelium was present, suggesting continual release of endothelium-derived relaxant factor (EDRF) at rest. When allowance was made for basal EDRF activity in aortic preparations, the maximal constrictor response to acetylcholine remained lower in the presence of endothelium, consistent with acetylcholine stimulation of EDRF, but maximal constrictor responses to the other three agents were the same with and without endothelium, suggesting that the direct constrictor response overrides EDRF activity.

Acetylcholine↗

The nature of endothelium-derived vascular relaxant factor.

The existence of endothelium-derived vascular relaxant factor (EDRF) was postulated by Furchgott and colleagues when they observed that acetylcholine paradoxically relaxed preconstricted aortic strip preparations by an endothelium-dependent mechanism. This phenomenon has since been demonstrated in different blood vessels and mammalian species and it can be elicited by several other agents. EDRF has been thought to be a humoral agent, a lipoxygenase derivative and possibly a free radical. In the study reported here, by using aortic preparations from the rabbit, alone and in cascade experiments with isolated perfused coronary preparations, we demonstrate definitively that EDRF is a humoral agent. It is released from unstimulated aortic preparations containing endothelium, its release can be stimulated for prolonged periods by acetylcholine, and it is not a lipoxygenase derivative or free radical but an unstable compound with a carbonyl group at or near its active site.

Acetylcholine↗

EDRF coordinates the behaviour of vascular resistance vessels.

Constriction of vascular smooth muscle in response to the stimulus of raised intravascular pressure--the myogenic response--represents a positive feedback mechanism which, if unopposed, could theoretically lead to instability in the intact circulation. Dilation in response to increased intraluminal flow would provide an opposing feedback mechanism which could confer overall stability. Flow-dependent dilation in conduit vessels is mediated by endothelium-derived relaxing factor (EDRF), but the relationship between flow and EDRF activity has not been studied in resistance vessels in situ. We here demonstrate that EDRF can coordinate the aggregate hydrodynamic properties of an intact network. Under control conditions, EDRF maintains a fourth-power relationship between diameter and flow so that the pressure gradient in each vessel asymptotically approaches a constant value at high flow rates. Basal EDRF release may also maintain a similar spatial distribution of flow at different flow rates, even under conditions of moderate pharmacological constriction.

Animals↗

Differences in basal endothelium-derived relaxing factor activity in different artery types.

Mechanical responses and calcium influx were measured in order to compare basal endothelium-derived relaxing factor (EDRF) activity in isolated preparations of rabbit aorta, rat aorta, and dog coronary artery. EDRF activity was characterized by endothelium-dependent mechanical relaxation and reduction of 45Ca influx which could be blocked by EDRF inhibitors. In resting preparations, the mechanical effects of basal EDRF were negligible in all preparations, and a small effect on calcium influx was demonstrated only in rat preparations. In agonist-constricted preparations, basal EDRF activity had only a small mechanical effect in rabbit preparations but markedly depressed constriction in rat and dog preparations; likewise, it had no demonstrable effect on calcium influx in rabbit preparations but had a marked effect in rat and dog preparations. In both resting and agonist-stimulated rabbit preparations, endothelium caused a cyclooxygenase product-dependent increase in calcium influx. Thus, basal EDRF activity has little or no effect in resting preparations and little or no effect in agonist-stimulated rabbit aorta preparations, but a marked effect in agonist-stimulated rat aorta and dog coronary preparations.

Animals↗

Nitric oxide-independent relaxations to acetylcholine and A23187 involve different routes of heterocellular communication. Role of Gap junctions and phospholipase A2.

NO- and prostanoid-independent relaxations are generally assumed to be mediated by an endothelium-derived hyperpolarizing factor (EDHF) that has been postulated to be an arachidonic acid metabolite. Recent evidence also suggests that direct heterocellular gap junctional communication (GJC) between endothelium and smooth muscle contributes to NO-independent relaxations. In the present study we have investigated the contribution of phospholipase A2 (PLA2)-linked metabolites and GJC to EDHF-type relaxations in rabbit mesenteric artery. In isolated rings preconstricted with 10 micromol/L phenylephrine in the presence of NG-nitro-L-arginine methyl ester (L-NAME) and indomethacin, acetylcholine (ACh) and the Ca2+ ionophore A23187 evoked relaxations that were markedly attenuated by the Ca2+-dependent PLA2 inhibitors 2-(p-amylcinnamoyl)amino-4-chlorobenzoic acid (3 micromol/L) and arachidonyl trifluoromethyl ketone (3 micromol/L), but were potentiated by the sulfhydryl agent thimerosal (300 nmol/L). In intact rings, relaxations to ACh were attenuated synergistically by L-NAME and Gap 27 peptide, an inhibitor of GJC, whereas ACh-evoked relaxations of "sandwich" preparations were unaffected by the peptide but were abolished by L-NAME. In both ring and sandwich preparations A23187-induced relaxations were attenuated by inhibition of PLA2 but were insensitive to L-NAME and Gap 27 peptide. We conclude that EDHF-type relaxations of rabbit mesenteric artery to ACh and A23187 depend on a common pathway that involves activation of PLA2. In the case of ACh, relaxation requires transfer of a factor or factors from the endothelium to smooth muscle via gap junctions, whereas A23187 permits release directly into the extracellular space.

8,11,14-Eicosatrienoic Acid↗

Mechanisms underlying chaotic vasomotion in isolated resistance arteries: roles of calcium and EDRF.

Nonlinear mathematical techniques now make it possible to quantify the complexity of an irregular time series through calculation of a parameter known as fractal dimension. In the present study, we use such an analysis to provide evidence that histamine-induced pressure oscillations in an isolated rabbit ear resistance artery are generated by deterministic rather than stochastic mechanisms, and that a minimum of 3 independent control variables is necessary to account for the complexity of the dynamics of these oscillations. The fractal dimension of the responses was independent both of the concentration of histamine used to induce rhythmic behavior, and the level of activity of the endogenous nitrovasodilator, EDRF. While both superficially influenced the form of the oscillations, it follows that neither are key control variables involved in their genesis. Nonlinear analysis of data obtained in the presence of NG-nitro-L-arginine methyl ester (L-NAME), which blocks EDRF synthesis, provided insights into the intrinsic smooth muscle control mechanisms responsible for generating rhythmic activity. The oscillations exhibited distinct "fast" and "slow" components (periods of 5-20 secs and 1-5 min. respectively). The former involved ion movements at the cell membrane and was inhibited by low [Ca2+]o, verapamil (which blocks voltage-dependent Ca2+ influx) and tetraethylammonium (which blocks Ca(2+)-activated outward K+ channels), whereas the latter involved Ca(2+)-induced Ca2+ release from intracellular stores and was inhibited by ryanodine. All such interventions decreased the overall fractal dimension of the responses to a value < 2, thus removing one degree of complexity (and hence control variable) from the dynamics. We conclude that the nonlinear interaction between a fast membrane oscillator and a slow intracellular oscillator generates chaos in vascular smooth muscle and that exogenous constrictor agonists and EDRF may be regarded as permissive and modulatory influences, respectively.

Animals↗