Contraction Of Intestinal Smooth Muscle Research Articles

Relations among canine intestinal motility, blood flow, and oxygenation.

Blood flow, arteriovenous O2 difference, and lumen pressure were measured in isolated loops of canine ileum. Ileal pressure was increased by an intra-arterial infusion of either Met-enkephalin or acetylcholine. Pressures were quantitated using a motility index (MI = mean of the pressure peaks divided by number of contractions per minute). Both Met-enkephalin and acetylcholine increased MI in a dose-dependent fashion. The highest MI achieved with acetylcholine was 37.9 mmHg, while Met-enkephalin produced a maximal MI of 8.1 mmHg. Ileal oxygen uptake increased when MI reached values greater than 6. There was a direct linear relationship between oxygen uptake and MI. Distension of the lumen in the absence of motility resulted in a decrease in oxygen uptake when lumen pressure reached 15-20 mmHg. The results of these studies indicate that contractions of intestinal smooth muscle can increase intestinal oxygen uptake and may contribute to the overall oxygen demands of the gut under conditions of fasting and feeding. Furthermore, large (greater than 20 mmHg) increments in lumen pressure during enhanced motility may compromise intestinal oxygenation.

Anamnestic stimulus-specific myoelectric responses associated with intestinal immunity in the rat

Changes in small intestinal motility reflective of local anaphylaxis are hypothesized to be associated with the rapid, immune-mediated rejection of infective larvae of the nematode Trichinella spiralis. This hypothesis was tested by comparing intestinal myoelectric activity in primarily and secondarily infected rats; the latter group represented immunized hosts, i.e., immunized by initial contact with the parasite. Patterns of slow waves, action potentials, interdigestive migrating myoelectric complexes, and migrating action-potential complexes induced by secondary infection differed from those associated with primary infection. Patterns in both cases differed from those in uninfected rats. Secondary but not primary infection evoked changes in myoelectric patterns within 15 min after administration of infective larvae. These changes could not be elicited by inoculation with dead larvae, larval excretory-secretory antigens, or a heterologous parasite, Eimeria nieschulzi. Results indicate that altered smooth muscle contractile activity is part of an anamnestic, stimulus-specific response to T. spiralis in immunized rats. These changes may represent the in vivo equivalent of the anaphylactically mediated intestinal smooth muscle contraction elicited in vitro in the Schultz-Dale reaction.

The binding of doxepin to histamine H1‐preceptors in guinea‐pig and rat brain

The affinity constant for doxepin obtained from inhibition of histamine-induced contraction of guinea-pig intestinal smooth muscle at 30 degrees C was 2.6 +/- 0.18 X 10(10)M-1. The slope of a Schild plot was not significantly different from unity. The affinity constant of doxepin did not vary markedly with temperature. At 37 degrees C it was 3.75 +/- 0.02 X 10(10)M-1 and at 25 degrees C 2.1 X 10(10)M-1. Doxepin was a competitive inhibitor of [3H]-mepyramine binding to guinea-pig cerebellar homogenates. The affinity constant derived for doxepin at 30 degrees C was 1.12 +/- 0.45 X 10(10)M-1. Hill coefficients for curves of doxepin or mepyramine inhibition of [3H]-mepyramine binding in guinea-pig cerebellum, cerebral cortex and hippocampus did not differ significantly from unity. The mean affinity of mepyramine for histamine H1-receptors in rat brain homogenates at 30 degrees C was 3.5 X 10(8)M-1. Hill coefficients for curves of doxepin or mepyramine inhibition of [3H]-mepyramine binding to homogenates of rat cerebral cortex or rat whole brain were near unity. These studies provide no evidence that doxepin binds preferentially to a sub-class of histamine H1-receptors in rat brain.

The presence in pig brain of an endogenous equivalent of apamin, the bee venom peptide that specifically blocks Ca2+-dependent K+ channels.

An apamin-like factor has been isolated from pig brain after extraction of the tissue and purification on sulfopropyl-Sephadex C-25 and on reversed-phase high pressure liquid chromatography. The apamin-like factor has the following properties: (i) it prevents 125I-labeled apamin binding to its specific receptor site present on rat brain synaptosomes, (ii) it is active in the radioimmunoassay for apamin (i.e., it prevents 125I-labeled apamin precipitation by anti-apamin antibodies), (iii) it induces contraction of guinea pig intestinal smooth muscle previously relaxed with epinephrine, and (iv) it blocks Ca2+-dependent K+ channels responsible for the long-lasting afterpotential hyperpolarization following the action potential in rat skeletal muscle cells in culture. All these properties are those of apamin itself. The apamin-like factor is a peptide that, like apamin, is destroyed by trypsin and unaffected by chymotrypsin. These results suggest the presence in mammalian brain of a potent Ca2+-dependent K+-channel modulator.

A synthetic peptide that is a bombesin receptor antagonist.

The tetradecapeptide bombesin was originally isolated from frog skin. Bombesin-like peptides have since been detected in mammalian gastrointestinal tract, brain and lung. These peptides have potent pharmacological effects on the central nervous system; they cause contraction of intestinal, uterine and urinary tract smooth muscle; and stimulate the release of other peptides including gastrin, cholecystokinin, motilin, pancreatic polypeptide, neurotensin, insulin, enteroglucagon, prolactin and growth hormone. Specific plasma membrane receptors for bombesin have been demonstrated on pancreatic acinar cells, brain membranes and pituitary cells. Studies defining the physiological importance of bombesin have been impeded by the lack of a bombesin receptor antagonist. Here we describe experiments which demonstrate that a peptide originally described as a substance P receptor antagonist, [D-Arg, D-Pro, D-Trp, Leu ]substance P, is also a bombesin receptor antagonist. This peptide competitively inhibits the ability of bombesin to stimulate enzyme secretion from dispersed pancreatic acini, and also inhibits the action of other peptides that interact with the bombesin receptor.

The inhibitory effect of X537A on vascular and intestinal smooth muscle contraction.

The effects of X537A (Lasalocid) on contractions induced in vascular and intestinal smooth muscles were examined. High K-induced sustained contractions were inhibited by X537A with an IC50 of 2.8 X 10(-6) M in rabbit aorta and 8.8 X 10(-8) M in guinea-pig Taenia coli. Changing the Ca concentration in the medium did not modify the effect of X537A. X537A also inhibited the noradrenaline-induced contraction in aorta (IC50 = 7.0 X 10(-8) M). In a solution without added glucose, the inhibitory effect of X537A on the K-induced contraction in aorta was augmented (IC50 = 1.5 X 10(-8) M). Under hypoxic conditions, the inhibitory effects of X537A on the noradrenaline-induced contractions in aorta and on the K-induced contraction in T. coli were decreased (IC50 greater than 10(-5) M and 3.2 X 10(-6) M, respectively). X537A inhibited the K-induced increase in cellular 45Ca content in aorta measured by a modified lanthanum method. However, the IC50 for 45Ca uptake (6.6 X 10(-7) M) was lower than that for K-induced contraction. In both tissues, X537A decreased the ATP content. Oxygen consumption of rat liver mitochondria was inhibited by X537A. From these results, it is concluded that the inhibitory effect of X537A on the noradrenaline-induced contraction in aorta and the K-induced contraction in T. coli, but not the K-induced contractions in aorta, may be due to the inhibition of aerobic metabolism.

The inhibitory effects of N2-dansyl-L-arginine-4-t-butylpiperidine amide (TI233) on contraction of vascular and intestinal smooth muscle.

Effects of N2-dansyl-L-arginine-4-t-butylpiperidine amide (TI233) on the contractions in vascular and intestinal smooth muscles were examined. High K-induced sustained contractions in the smooth muscles were inhibited by TI233 with an IC50 of 2.1 X 10(-5) M for rabbit aorta and 3.6 X 10(-6) M for guinea-pig taenia coli in a solution containing 1.5 mM Ca. Initial transient contraction induced by K in taenia coli was less sensitive to the inhibitory effect of TI233. When the Ca concentration in the medium was decreased to 0.3 mM, the concentration-inhibition curves for TI233 shifted to the left in both aorta and taenia coli. Increasing the Ca concentration to 7.5 mM shifted the curve to the right in the aorta. TI233 also inhibited the noradrenaline-induced contraction in the aorta (IC50 = 2.1 X 10(-5 M). In a hypoxic solution without added glucose, the inhibitory effect of TI233 on the K-induced contraction in aorta was augmented. In the presence of high concentrations (40 mM) of glucose in hypoxia, TI233 did not inhibit the noradrenaline-induced contraction of the aorta. Hypoxia and a high concentration of glucose also decreased the inhibitory effect of TI233 on the K-induced contraction in taenia coli. TI233 inhibited the K-induced increase in cellular Ca content measured by a modified lanthanum method. TI233 decreased oxygen consumption and ATP content of resting and K-stimulated aorta and taenia coli. It was concluded that TI233 inhibits the vascular and intestinal smooth muscle contraction by a Ca antagonistic action and also by inhibition of aerobic metabolism.

Darmstoff analogues. 2. Ring and side-chain effects on smooth-muscle contraction.

2-cis-delta 8-Heptadecenyl-4-(hydroxymethyl)-1,3-dioxolane monosodium phosphate (1b) has been shown to be present as a major component of Darmstoff in mammalian intestine and to be a potent inducer for contraction of intestinal smooth muscle. The analogous 2-pentadecyl material 1a, also found abundantly in the intestine, is inactive. In this study, synthesis of phosphorylated hydroxymethyl-1,4-dioxanes, -tetrahydrofurans, -cyclopentanes, and -oxathiolanes bearing both oleyl and palmityl side chains is reported. Of these, 2-(hydroxymethyl)-5-cis-delta 8-heptadecenyl-1,4-dioxane monosodium phosphate (2b) exhibits about 12% of the activity of 1b. Its pentadecyl analogue 2a, like 1a, is totally inactive, as are all other compounds prepared. The results indicate that Darmstoff-like compounds exhibit specific chemical requirements for activity and that where activity is encountered, the side-chain specificity noted in 1a and 1b can be preserved.

Tris does not inhibit isolated vascular or intestinal smooth muscle contraction.

There are conflicting reports on the effect of tris(hydroxymethyl)aminomethane (Tris) on smooth muscle contraction. Therefore, the effect of substitution of Tris for bicarbonate in physiological saline solution on smooth muscle contractility was investigated. Tris (25 mM) increased the amplitude without changing the frequency of spontaneous contractions in rat portal vein. Tris did not inhibit either norepinephrine-induced contractions in rat portal vein and aorta or histamine-induced contractions in taenia coli. When the pH of the Tris-buffered solution was lowered from the control value of 7.4 to 7.0, spontaneous contractions in portal vein as well as agonist-induced contractions in all of the above preparations were inhibited. Tris did not inhibit contractions induced by calcium in potassium-depolarized smooth muscle preparations if the Tris-buffered solution contained sufficient amounts of sodium ion (as did bicarbonate-buffered solution). These results suggest that Tris does not inhibit contractile responses of isolated vascular and intestinal smooth muscle preparations provided that the conditions other than the buffer system (especially pH of solution) are similar to those in bicarbonate-buffered solutions.

Inhibition of Intestinal Smooth Muscle Contraction by Surface‐active Alkyltrimethylammonium Salts

The effects of surface-active alkyltrimethylammonium salts (C10--C20) on the contractile activity of isolated rat jejunum and on the gastrointestinal motility in anaesthetized rats were studied. The surfactants caused a dose-dependent depression of spontaneous contractions and of acetyl-beta-methylcholine and BaCl2 induced contractions of isolated rat jejunum. The depressant activity of the surface-active alkyltrimethylammonium salts on smooth muscle contraction increased with an increase in the length of the alkyl chain to maximum activity at C16. It is suggested that the depressant activity of the surface-active alkyltrimethylammonium salts on smooth muscle contraction is due to a non-specific interaction of the surfactants with the cellmembrane of the muscle cells. No effect on gastric and intestinal motility in vivo was observed following intragastric or intraintestinal administrations of surface-active alkyltrimethylammonium salts. This lack of effect upon intraintestinal or intragastric administration probably depends on an inability of the surfactants to reach the site of action when administered from the mucosal side.

Contractions of cat small intestinal smooth muscle in calcium-free solution.

A rise in internal Ca2+ is the signal which activates the contractile mechanism in muscle. In striated muscle there is a well developed sarcoplasmic reticulum which stores and releases the bulk of the Ca2+ involved in the contraction cycle1,2, allowing the muscle to operate relatively independently of external calcium levels. In smooth muscle the sarcoplasmic reticulum system is much less extensive3,4 and normal contractile activity is very sensitive to calcium levels in the bathing medium5–7. These findings, together with the small size of smooth muscle cells which minimises diffusion time from the plasma membrane to the contractile apparatus, have suggested to some investigators that the Ca2+ which enters the cell during Ca2+-mediated action potentials is either sufficient by itself to activate contraction or is a necessary trigger for the further release of Ca2+ from internal stores8–11. This notion is given support by the observation that contractile activity rapidly dies out when smooth muscle preparations are exposed to calcium-free solutions. However, electrical activity is eventually lost in these solutions. We report here on spontaneous contractile and electrical activity, and evoked mechanical activity, recorded after prolonged exposure to calcium-free EGTA saline in intestinal smooth muscle of cat.

The effects of theophylline and cyclic AMP on intestinal smooth muscle contractions

The effects of 10 −4-10 −3 M theopphylline, cyclic 3'5'-adenosine monophosphate (cyclic AMP), dibutyryl cyclic AMP (DBC) and theophylline plus DBC were observed on the spontaneous and potassium (K)-induced contractions of the guinea-pig taenia coli. All of the compounds inhibited spontaneous contractions as well as the phasic and tonic components of the K-contracture. The effects of cyclic AMP, a compound which will not readily penetrate the cell membrane, were similar to those of the other compounds in inhibiting the spontaneous and phasic contractions, but it was a less effective inhibitor of the tonic contracture. Inhibition of spontaneous and phasic activity may occur at the cell membrane while the inhibitory actions on the tonic contracture may depend on cellular penetration by the inhibitor. There was no evidence that theophylline and DBC have different sites of action.

Relationship between cyclic AMP, phosphodiesterase activity, calcium and contraction in intestinal smooth muscle.

AbstractEffects of contracting agents on the mechanical and the metabolic activity in smooth muscle have been studied. The contraction induced by carbacholine in colon muscle from rabbit was preceded by a decrease of the cyclic AMP content. After development of the tension, the level of cyclic AMP was increased, the phosphodiesterase activity was reduced, and the phosphorylase SaL activity and glycogenosis were stimulated. These effects were blocked by atropine but not by an adrenergic β‐blocking agent. In the Ca‐poor muscle there was a sustained decrease of the cyclic AMP content by carbacholine. Contraction induced by K+ ions showed similar metabolic actions as carbacholine and these effects were Ca++ dependent too. The contracting effect of imidazole was preceded by an activation of phosphodiesterase and by reduction of cyclic AMP. The metabolic effects induced by imidazole were not Ca++ dependent. Imidazole potentiated the contracting action of carbacholine and K+. Ca++ in the same concentrations as those activating the contractile system, inhibited the phosphodiesterase activity in homogenate of colon muscle. The relationship between cyclic AMP, Ca++ and contraction in smooth muscle is discussed.

Role of muscarinic cholinergic receptors in regulation of guanosine 3':5'-cyclic monophosphate content in mammalian brain, heart muscle, and intestinal smooth muscle.

Effects of acetylcholine and of agents that mimic or block its physiological actions have been studied upon concentrations of guanosine 3':5'-cyclic monophosphate (cyclic GMP) and adenosine 3':5'-cyclic monophosphate (cyclic AMP) in slices of mammalian cerebral cortex, heart ventricle, and ileum. Acetylcholine, and cholinomimetic agents with a predominantly muscarinic action, such as methacholine, bethanechol, and pilocarpine, induced an increase in the concentration of cyclic GMP, accompanied by no change or a slight decrease in the concentration of cyclic AMP, in all three tissues studied. Tetramethylammonium, a cholinomimetic agent with a predominantly nicotinic action, on the other hand, did not significantly alter concentrations of either cyclic GMP or cyclic AMP. The increase in the tissue content of cyclic GMP induced by acetylcholine and its muscarinic analogs was antagonized by atropine, a muscarinic blocking agent, but not by hexamethonium, a nicotinic blocking agent. These and other results suggest the generalization that the interaction of acetylcholine with muscarinic receptors, but not with nicotinic receptors, causes an increase in the concentration of cyclic GMP. The data are compatible with the hypothesis that cyclic GMP may mediate the muscarinic actions of acetylcholine. The antagonistic actions of cholinergic and adrenergic agents on the physiology of contraction of intestinal smooth muscle and of cardiac muscle are reflected in, and may result from, antagonistic actions of these compounds on the levels of both cyclic GMP and cyclic AMP in these tissues.

The effect of catecholamine infusions on myocardial blood flow, metabolic heat production and on general haemodynamics, before and after alprenolol (H56/28), in anaesthetized cats

1. In cats anaesthetized with pentobarbitone sodium, infusions of adrenaline, noradrenaline (0.5 mug/kg per min) and isoprenaline (0.25 mug/kg per min) increased myocardial blood flow, myocardial heat production, left ventricular systolic and end-diastolic pressures, left ventricular +ve and -ve dp/dt max, and calculated cardiac output, effort and oxygen consumption. These effects (apart from the effect of noradrenaline on left ventricular systolic pressure) were markedly reduced by previous administration of alprenolol (0.5 or 1.0 mg/kg).2. Infusions of adrenaline and noradrenaline increased arterial diastolic blood pressure and isoprenaline reduced it. After alprenolol the effects of adrenaline and noradrenaline were potentiated and that of isoprenaline abolished; in some experiments isoprenaline increased arterial diastolic pressure after alprenolol. Alprenolol did not influence the increases in arterial systolic pressure which followed the administration of adrenaline and noradrenaline.3. Isoprenaline-induced tachycardia was markedly reduced and adrenaline tachycardia was converted to bradycardia after alprenolol. The bradycardia which occurred during noradrenaline infusions was unaffected.4. After blockade by alprenolol, recovery of the effects of isoprenaline on left ventricular dp/dt and on heart rate occurred more quickly than recovery of the effects on arterial diastolic pressure. This suggests that alprenolol has a greater affinity for beta(2)- than for beta(1)-adrenoceptors.5. Intravenous administration of acetylcholine decreased arterial blood pressure, left ventricular pressure and +ve and -ve dp/dt max. During recovery from these effects there was a marked increase in +ve dp/dt max. which was absent after the administration of alprenolol (0.5 mg/kg). Because this dose of alprenolol is thus able to block the effects of reflex sympathetic cardiac nerve stimulation but does not completely antagonize the effects of exogenous adrenaline on dp/dt, it is suggested that alprenolol may have some adrenergic neurone blocking activity.6. Increases in liver and myocardial blood flow and heat production produced by noradrenaline, adrenaline and isoprenaline were reduced after alprenolol.7. Isoprenaline reduced air-way resistance and this effect was abolished by alprenolol; increases in air-way resistance produced by adrenaline and nor-adrenaline were augmented. All three amines inhibited intestinal smooth muscle contractions in vivo. Only the effect of isoprenaline was reduced by alprenolol.

Mobilization of cellular calcium for contraction in intestinal smooth muscle.

Mobilization of cellular calcium for contraction in intestinal smooth muscle.

Effect of metoclopramide on transport in the small intestine of the dog.

A method is described for the simultaneous measurement of flow rate, mean transit time, and volume of the intestine during the transport of fluid through a segment of the small intestine of the dog. Using this method, the effect of metoclopramide on motility of the small bowel was assessed. The drug resulted in a decrease in transit time and volume in the test segment of bowel while the flow rate remained constant. This finding suggests that metoclopramide-stimulated contraction of intestinal smooth muscle.

Hypolipidemic agents.

Despite the fact that the role of lipids and cholesterol in atherosclerosis has not been totally clarified to the clinician's satisfaction, medicinal chemistry has been very successful in this last decade in showing that a diverse group of compounds is capable of reducing blood lipid levels in many animal species and in humans. The pharmacological basis for the evaluation of many hypolipidemic agents is summarized. The prostaglandins are vasodepressor agents. They inhibit uterine contraction, yet, on the other hand, they increase cardiac output and induce contraction of intestinal smooth muscle. Prostaglandins exhibit marked antilipolytic activity. It has been shown that 0.2 mug/kg/min, of intravenous infusion of prostaglandin E1 produced an increase in serum-free fatty acide levels in the dog. At a 4 times higher rate of infusion, the plasma-free fatty acid concentration was lowered. The pharmacological activities of individual prostaglandins embrace a great variety of biological phenomena. In certain systems their effects are antagonistic. The rationale for the use of estrogens as hypolipidemics is the observation of the low incidence of atherosclerotic heart disease in Caucasian women of childbearing age.

Acetylcholine and calcium on membrane permeability and contraction of intestinal smooth muscle.

Acetylcholine elicited a sustained contraction and an increase in potassium efflux in longitudinal muscle isolated from the guinea pig ileum. Stepwise increases in the calcium concentration of the bathing medium, from 0.06 to 36 mM generally reduced the increase in potassium efflux, but had a complex effect on the mechanical response. Contractions produced by high levels of acetylcholine became progressively larger or remained at a high magnitude as the calcium concentration was increased. Contractions produced by low levels of acetylcholine also improved initially, but were depressed again by the highest concentration of calcium introduced. Ethanol, in the appropriate concentration, inhibited completely the acetylcholine-induced contraction without reducing the increase in potassium efflux. Calcium reversed this effect. Both extracellular calcium and ethanol depressed the large, transient increase in muscle tone developed by fibers that were preincubated in a high calcium medium and then exposed to a calcium-free medium. These findings suggested that extracellular calcium ions react with two different sites in the membrane, a stabilizing site and a storage site. A muscle contraction is activated by calcium ions which diffuse from the storage site to the myoplasm. Calcium ions reacting with the stabilizing site impede this diffusion process. Part of the stimulatory effect of acetylcholine is derived from its capacity to counteract the action of calcium at the stabilizing site.

Blockade of the biochemical correlates of contraction and relaxation in uterine and intestinal smooth muscle.

The relationship between contraction and glycogen phosphorylase activation by adrenergic agents has been studied extensively in cardiac and skeletal muscle (see review by Haugaard & Hess, 1965). I t was suggested by Bulbring in 1960 that smooth muscle relaxation induced by epinephrine was a process requiring increased metabolic energy; and early results (Axelsson et al . , 1961) indicated tha t the action of epinephrine on the smooth muscle cell membrane might be related to its ability t o increase phosphorylase a activity. However, later reports concluded that the two effects were not related (Timms et al., 1962) and that, in fact, the increases in phosphorylase a activity seen in previous experiments were artifactual (Bueding et al., 1962). The present study is concerned with the effects of catecholamines and adrenergic blocking agents on the motility and phosphorylase activity of isolated rat uterus and guinea pig taenia coli.