Tamoxifen induces vasorelaxation via inhibition of mitogen-activated protein kinase in rat aortic smooth muscle

The contribution of the mitogen-activated protein kinase (MAPK) pathway to the relaxation induced by tamoxifen, a synthetic non-steroidal anti-estrogen, was examined in rat vascular smooth muscle. Tamoxifen (0.1-300 microM) inhibited the contraction induced by endothelin-1 (ET-1, 3 nM) in aortic smooth muscle in a concentration-dependent manner. The inhibitory effect of tamoxifen was not attenuated by 10 microM ICI 182,780, a selective antagonist of estrogen receptors. In the Ca(2+) channel inhibitor verapamil (1 microM)-pretreated strips, tamoxifen also inhibited the contraction induced by ET-1. Both PD098059 and SB203580, inhibitors of MAPK/extracellular signal-regulated kinase (ERK) kinase and p38 MAPK, respectively, inhibited ET-1-induced contraction in aortic smooth muscle. In Western blot analysis with anti-phosphorylated MAPK antibodies, ET-1 (3 nM) enhanced activities of both ERK1/2 and p38 MAPK in aortic muscle strips, which were not attenuated by the treatment with 4 mM EGTA. Tamoxifen (100 microM) inhibited the activities of ERK1/2 and p38 MAPK induced by ET-1 without significant changes in the expression of these kinases. These results suggest that tamoxifen induces relaxation of rat vascular smooth muscle, and that this is, at least in part, mediated by the inhibition of the Ca(2+)-independent MAPK pathway.     

Palmar digital vessel relaxation in healthy horses and in horses given carbohydrate

OBJECTIVE: To compare in vitro smooth muscle relaxation of palmar digital vessels from healthy horses with those from horses in the prodromal stage of experimentally (carbohydrate) induced laminitis. ANIMALS: 16 adult horses. PROCEDURE: Segments of palmar digital vessels were obtained from 5 healthy horses and 6 horses given carbohydrate. Vascular rings from the palmar digital artery and vein were suspended in individual organ baths containing buffer solution and indomethacin; isometric tension was recorded, and contraction and relaxation were compared. Smooth muscle contraction in response to cumulative addition of phenylephrine was recorded in the absence and presence of 1 microM NG-nitro-L-arginine methyl ester (L -NAME). After wash out, vascular rings were preconstricted with phenylephrine (0.3 microM), and cumulative endothelium-dependent (acetylcholine-induced) and independent (nitroprusside-induced) smooth muscle relaxations were recorded in the absence or presence of L -NAME. RESULTS: Phenylephrine increased vascular smooth muscle tone in ring preparations of palmar digital arteries and veins. Addition of acetylcholine or nitroprusside induced relaxation of palmar digital artery and vein ring preparations. Use of L-NAME (1 microM) significantly reduced maximal relaxation induced by acetylcholine, but not by nitroprusside. Maximal relaxation induced by acetylcholine, but not by nitroprusside, was reduced in vascular rings prepared from carbohydrate-overloaded horses. CONCLUSION AND CLINICAL RELEVANCE: Reduced endothelium-dependent relaxation of palmar digital vessels may have a role in the pathophysiology of acute laminitis after carbohydrate overload in horses.     

Arterial supply of the penis in the New Zealand rabbit (Oryctolagus cuniculus L.)

In the present study, the distributional pattern of the penile artery and the vessels joining the blood supply of the penis were investigated in the New Zealand rabbit. Eight adult rabbits were used in the study. In order to exhibit the vascular network by dissection, latex was injected via the abdominal aorta. The main vessel which supplies blood to the penis, the penile artery, is a branch of the internal pudendal artery. It divides into two branches which form the deep and dorsal penile arteries at the level of the ischiadic arch. The deep penile artery penetrates the tunica albuginea, and forms the arterial network of corpus cavernosum penis. On the other hand, the dorsal penile artery gives off three small branches for the subischiocavernosus muscle and at the level of the attachment of this muscle sends two small branches for the preputium. The course of both arteries follows the dorsolateral surface of the penis to the glans and ends in an anastomosis. Hence, a caudal branch of the prostatic artery which originates from the umbilical artery joins the blood supply of the penis in the rabbit. After vascularizing the prostate complex, it ends by entering the corpus spongiosus penis at the dorsolateral surface at the level of the ischiadic arch.     

Effects and bioassay of Escherichia coli enterotoxin (heat-stable fraction) on smooth muscle

Heat-stable E. coli and V. cholerae enterotoxins and E. coli endotoxin were tested on the following smooth muscle preparation: vascular; rabbit aorta; rat mesenteric arterioles, and intestinal, rabbit jejunum; pig duodenum; dog jejunal lamina propria smooth muscle. The results indicated that in most preparations used, the prime action of heat-stable enterotoxin from pathogenic strains of E. coli consisted in neutralizing the effects of both alpha or beta adrenergic agonists. In this respect the effect of enterotoxin appeared similar to that of alpha or beta adrenergic blockers. Using the same models, it was found that this enterotoxin did not interfere with the effects of cholinergic agonists or of biogenic amines. Control heat-stable enterotoxin preparations and other tested substances proved inactive, suggesting that different receptor sites might exist for these agents in our models. It appears that smooth muscle preparations are well suited for bioassay of active heat-stable E. coli enterotoxin.     

Failure of 3-methylindole to contract the bovine pulmonary vein or to release mediators of anaphylaxis from the bovine lung in vitro

Strips of smooth muscle from the pulmonary vein, pulmonary artery, trachea and bronchus of calves were incubated in an organ bath with 3-methylindole (3MI) and 3-methyloxindole (3MOI). 3MI and 3MOI (5-640 micrograms/ml) did not cause contraction of any of the isolated smooth muscle preparations. No evidence for the release of mediators of anaphylaxis was obtained when chopped bovine lung preparations were incubated with 3MI (20 micrograms/ml) and 3MOI (25 micrograms/ml). Results of the present work diminish the possibility that the pneumotoxic effect of 3MI is due to a primary hydrodynamic imbalance across the alveolocapillary membrane resulting in excess filtration over reabsorption.     

Aberrant right subclavian artery causing megaoesophagus in three cats

Three entire, domestic, shorthair male cats (age range: 3 months to 5 years) were referred because of regurgitation. Megaoesophagus attributable to aberrant right subclavian artery, originating from the aorta at the level of the fourth intercostal space, was diagnosed in all cats using thoracic radiography and CT angiography. One cat had concurrent patent ductus arteriosus with a normal aortic arch. Three‐dimensional volume‐rendered CT images were used to assess the malformations and to plan surgery for the treatment of the vascular anomalies. Different surgical approaches were used in the two kittens. The third cat was not operated. CT angiography is well suited for preoperative planning in cats with aberrant right subclavian artery alone or in combination with other vascular anomalies.     

Familial aortic aneurysm in Leonberg dogs

A thoracic aortic aneurysm was diagnosed in a 6-month-old male Leonberg dog by use of radiography, transthoracic and transesophageal echocardiography, and magnetic resonance imaging. The aneurysm was associated with a twisted ascending aorta and dilatation of several other thoracic arteries (pulmonary trunk, brachiocephalic trunk, and left subclavian artery). Histologic examination of the aorta revealed cystic medial necrosis, with disruption of the elastic network, collagen fibers, and the muscle glycoprotein fibrillin-1. The dam and sire of the dog and 8 littermates were examined by use of transthoracic echocardiography. The sire and 1 male littermate also had an aneurysm of the ascending aorta. To the authors' knowledge, this is the first report of familial aortic aneurysm in dogs.     

Disturbances in ex vivo vascular smooth muscle responses following exposure to Pasteurella haemolytica vaccines

Weekley, L.B., Eyre, P. Disturbances in ex vivo vascular smooth muscle responses following exposure to Pasturella haemolytica vaccines. J. vet. Pharmacol. Therap. 16 , 446–453.
Rats were vaccinated with saline (control) or one of the two commercially available Pasteurella haemolytica vaccines Presponse or Precon-PH. Animals were killed 3 days later and thoracic aorta removed for evaluation of the ex vivo biophysical responses to carbachol (CCh). In some experiments, vascular endothelium was mechanically removed. Vaccination of rats impairs the endothelial-dependent relaxation to CCh. In vessels with endothelium removed, the contractile response to CCh is converted into a relaxation following vaccination. Treatment of endothelial-denuded vascular rings ex vivo with methylene blue, a guanylate cyclase inhibitor, reduced the vaccination effect. Treatment of vascular rings with the superoxide dismutase inhibitor diethyl-dithiocarbamate, impairs the relaxant reponse of de-endothelialized vessels to CCh in Presponse vaccinated rats while enhancing the relaxation response of vessels from Precon-PH vaccinated rats. De-endothelialized vessels from vaccinated rats, but not control rats, relaxed in the presence of 7V-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide synthetase. Furthermore, in the presence of L-NMMA, the relaxant response to CCh is significantly enhanced by Precon-PH but not Presponse. The normal relaxant response to hydrogen peroxide is converted into a contraction following vaccination. Results suggest that exposure to commercially available P. haemolytica vaccines alters vascular smooth muscle reactivity to CCh and that several independent pathways may be altered.     

Computed tomography angiography of coarctation of the aorta in a dog

Coarctation of the aorta has been described previously as either a post-mortem or angiographic finding in three dogs with clinical signs related to the aortic coarctation. A 10-year-old dog was presented for evaluation of suspected laryngeal paralysis. On physical examination, femoral pulses were absent bilaterally, with an indirect systolic blood pressure difference of 60 mmHg between the ipsilateral thoracic and pelvic limbs. Coarctation of the aorta was detected on a thoracic computed tomographic angiography study. The coarctation was pre-ductal in position, with extensive dilation of the descending thoracic aorta. Characteristic rib changes that are seen in humans with coarctation were not apparent in this dog. 3D reconstructions of the thorax provided high vascular definition with exact localization of the aortic narrowing.     

Histamine receptors mediate contraction of reticular groove smooth muscle of adult goats

The present study was conducted to evaluate the effect of histamine and to characterise its receptor subtypes in reticular groove (RG) smooth muscle of adult goats. The studies were done using floor and lip regions of RG. We used tension experiments on smooth muscle of RG isolated from adult goat for functional characterisation of H₁ and H₂ receptors. Western blotting and immunohistochemistry experiments were conducted for molecular characterisation of these receptors. Histamine evoked concentration-dependent contraction of isolated RG circular and longitudinal smooth muscle preparation. Pyrilamine antagonised the action of histamine. Histamine did not induce any relaxant effect on RG preparations. Additionally, cimetidine did not produce any significant effect on histamine-induced response. Non-selective histaminic receptor antagonist cyproheptadine attenuated the contraction response to histamine in the smooth muscle. Molecular characterisation and localisation of H₁ and H₂ receptor proteins confirmed the presence of these receptors in RG. It is most likely that histamine-induced contractile effect in RG smooth muscle of goats is mediated by H₁ histaminic receptors.     

CD38 gene disruption inhibits the contraction induced by alpha-adrenoceptor stimulation in mouse aorta

CD38 is an ectoenzyme with ADP-ribosyl cyclase and hydrolase activities, which synthesizes cyclic ADP-ribose from NAD and hydrolyzes cyclic ADP-ribose to ADP-ribose. It has been shown that cyclic ADP-ribose is a potent Ca(2+) mobilizing messenger in many cells. To know the physiological role of cyclic ADP-ribose in vascular smooth muscle, we examined the effects of various agonists in the aorta isolated from CD38 knockout (CD38(-/-)) mouse. Western blot analysis showed that CD38 protein was detected in the aorta isolated from wild-type (CD38(+/+)) mouse, but not from CD38(-/-) mouse. In the aortae isolated from both CD38(+/+) and CD38(-/-) mice, KCl, phenylephrine and norepinephrine induced concentration-dependent contraction. KCl produced similar concentration-dependent responses in the aortae from both CD38(+/+) and CD38(-/-) mice. Maximum force of contraction induced by KCl (65 mM) was same in the size. Phenylephrine- and norepinephrine-induced contractions were, however, significantly smaller in the aortae from CD38(-/-) mice than in those from CD38(+/+) mice. 5-Hydroxytryptamine, endothelin-1, caffeine and thapsigargin-induced contractions were not significantly different in these two aortae. These results suggest that CD38 gene disruption inhibits alpha-adrenoceptor-induced vascular contractions and cyclic ADP-ribose-mediated signal transduction system is committed in these responses.     

Smooth muscle inhibition by an extract of Sesbania drummondii

The acute effects of an extract of Sesbania drummondii were assessed in vitro on the smooth muscle contractility of intestine and lung parenchyma in the chicken and aortic arch in the rat. Dose-response contraction curves for histamine, carbachol, and norepinephrine were obtained in the ileal, parenchymal, and aortic strips, respectively. After washing was completed, the strips returned to baseline tensions and then were incubated for 10 minutes with an ethyl acetate extraction fraction of S drummondii (molecular weight less than 500). Dose-response curves to the same agonists were repeated at the end of the incubation period. Contractile responsiveness of chicken ileum was little affected by acute incubation of the tissue with the extract. Comparison with the great inhibition of in vitro ileal contractility seen previously in chickens with chronic toxicosis indicated that intestinal inhibition was not due to acute effects of sesbania, but required time for a toxic metabolite to be formed or for damage to occur from affected vasculature. Contractile responsiveness of chicken lung parenchyma to histamine (10(-5) M and 10(-4) M) was significantly decreased, as was rat aortic responsiveness to norepinephrine (10(-7) M to 10(-4) M). Responses in parenchyma were not as greatly inhibited as those in tissue from animals with chronic toxicosis. Greatest inhibition of contractility was seen in the vascular strips, indicating that vascular inhibition has a role in pathologic changes. To test the vasculature inhibition effect in vivo, anesthetized, catheterized rats were given 100-microliter aliquots of dilutions of the extract, IV.(ABSTRACT TRUNCATED AT 250 WORDS)     

Aortic rupture in poultry: a review

Mortality in poultry due to aortic rupture is characterized by sudden death. The condition is seen in fast-growing male turkeys but has also been described in chickens, ostriches, and waterfowl. Losses in affected flocks usually only reach 1-2%. Post-mortem examination shows a large blood clot in the abdominal cavity subsequent to a dissecting aneurysm. Fragmentation of elastic fibres and degenerative changes of smooth muscle cells are seen in the region of the rupture. Intimal sclerotic plaques are present adjacent to the site of rupture. Copper deficiency, hypertension, hormonal influences, diet, lathyrism, zinc deficiency, pharmaceuticals, and parasites are precipitating factors for aortic rupture. Field studies suggest that favourable results are obtained with reserpine and copper as treatment for ruptured aorta.     

Possible involvement of K+ channel opening to the interleukin-1 beta-induced inhibition of vascular smooth muscle contraction.

We have previously shown that interleukin-1 beta relaxes vascular smooth muscle by the NO-dependent and independent mechanisms (Takizawa et al.: Eur. J. Pharmacol. 330: 143-150, 1997). In this study, we investigated the mechanism of NO-independent relaxation. Treatment of the rat aorta with interleukin-1 beta for 24 hr inhibited the high-K+ induced contraction by decreasing cytosolic Ca2+ level ([Ca2+]i). The relationship between [Ca2+]i and tension in intact muscle and the pCa-tension curves in permeabilized muscle suggested that Ca2+ sensitivity of contractile element was not changed after the interleukin-1 beta-treatment. After a treatment with interleukin-1 beta for 24 hr, contractile effects of phenylephrine (1 microM-10 microM) were markedly inhibited in the presence of L-NMMA (100 microM) applied to inhibit NO synthesis. A blocker of ATP-sensitive K+ channel, glibenclamide (1 microM), partially recovered the interleukin-1 beta-induced inhibition. In contrast, a blocker of Ca(2+)-activated K+ channel, charybdotoxin (0.1 microM), was ineffective. These results suggest that membrane hyperpolarization due to activation of ATP-sensitive K+ channels may partly be responsible for the NO-independent mechanism of interleukin-1 beta-induced inhibition of vascular smooth muscle contraction.     

Effects of xylazine on canine coronary artery vascular rings

OBJECTIVE: To determine the effects of xylazine on canine coronary artery smooth muscle tone. SAMPLE POPULATION: Hearts of 26 healthy dogs. PROCEDURE: Dogs were anesthetized with pentobarbital, and vascular rings of various diameters were prepared from the epicardial coronary arteries. Vascular rings were placed in tissue baths to which xylazine was added (cumulative concentrations ranging from 10(-10) to 10(-4) M), and changes in vascular ring tension were continuously recorded. Effects of the nitric oxide inhibitor NG-nitro-L-arginine methyl ester (L NAME; 5 mM), the alpha1-adrenoceptor antagonist prazosin (10 mM), and the alpha2-adrenoceptor antagonist atipamezole (10 mM) on xylazine-induced changes in vascular ring tension were determined. Results were expressed as percentage of maximal contraction for each vascular ring preparation. RESULTS: Xylazine induced vasoconstriction of small (< 500-microm-diameter) and medium (500- to 1,000-microm-diameter) vascular rings but not of large (> 1,000-microm-diameter) rings. For large vascular rings, L-NAME, atipamezole, and prazosin did not significantly affect the contractile response to xylazine. For small vascular rings, the contractile response following addition of xylazine to rings treated with L-NAME was not significantly different from the contractile response following addition of xylazine to control rings, except at a xylazine concentration of 10(-6) M. Xylazine-induced vasoconstriction of small vascular rings was blocked by atipamezole, but the addition of prazosin had no effect on xylazine-induced vasoconstriction. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that xylazine increases smooth muscle tone of small canine coronary arteriesand that this effect is predominantly mediated by stimulation of alpha2adrenoceptors.     

Internal thoracic artery-caudal epigastric artery as a collateral pathway in a dog with aortic occlusion: a case report

Aortic occlusion is a rare vascular disorder in the dog. This report describes the clinical feature of an aged Boxer with complete occlusion of the abdominal aorta caused by an intraluminal thrombus. Angiography evidenced a collateral circulation, represented by the internal thoracic artery, the cranial epigastric artery and the caudal epigastric artery, providing adequate blood flow to the pelvis and pelvic limbs. This vascular network has only recently been recognised as one of the major collateral pathways for arterial blood supply to the pelvis and lower extremities in humans with chronic aorto-iliac occlusive disease (CAOD). Furthermore, a femoral artery Doppler waveform, characterised by low amplitude, a monophasic systolic wave with blunted parabolic flow profile and a small spectral window, peculiar to humans with CAOD, is documented for the first time in a dog with aortic occlusion.     

Effect of pharmacological agonists on contractile responses in aortic rings derived from endotoxaemic rats

To investigate the vascular smooth muscle dysfunction of septic shock, in vitro isometric responses to phenylephrine (PE) and acetylcholine (ACh) were evaluated in aortic rings, with and without endothelium (± E), removed from male Wistar rats 1.5, 3 and 6 h after intravenous (i.v.) administration of 5 mg/kg lipopolysaccharide (LPS) or vehicle. A reduction in maximum contraction (±E) and sensitivity (-E) to PE were identified at 6 but not at 1.5 or 3 h. Maximum relaxation to ACh (+ E) was not affected by LPS treatment but sensitivity was increased at 1.5 and 3 h. Having identified 6 h as the time at which the most pronounced changes were observed, further studies at this interval found that maximum contraction to potassium chloride (±E), prostaglandin F_2a (+ E) and detomidine (-E) and relaxation to salbutamol (-E) were less in aortic rings from endotoxaemic rats. Sensitivity to KCl (±E), PGF_2a (- E) and detomidine (- E) was also reduced. Relaxation to sodium nitroprusside and atrial natriuretic peptide was not changed. These results suggest that attenuated pressor responses to a variety of vasoactive agents may be expected in patients 6 h after systemic exposure to endotoxin and that this vasoplegia may influence the vascular side-effects of therapeutic agents.     

Failure of 3-methylindole to contract the bovine pulmonary vein or to release mediators of anaphylaxis from the bovine lung in vitro

Strips of smooth muscle from the pulmonary vein, pulmonary artery, trachea and bronchus of calves were incubated in an organ bath with 3-methylindole (3MI) and 3-methyloxindole (3MOI). 3MI and 3MOI (5–640 g/ml) did not cause contraction of any of the isolated smooth muscle preparations. No evidence for the release of mediators of anaphylaxis was obtained when chopped bovine lung preparations were incubated with 3MI (20 g/ml) and 3MOI (25 g/ml). Results of the present work diminish the possibility that the pneumotoxic effect of 3MI is due to a primary hydrodynamic imbalance across the alveolocapillary membrane resulting in excess filtration over reabsorption.     

Unusual vascular ring anomaly associated with a persistent right aortic arch in two dogs

An unusual vascular ring anomaly consisting of a persistent right aortic arch and a left ligamentum arteriosum extending from the main pulmonary artery to an aberrant left subclavian artery and left aortic arch remnant complex was identified in a German shepherd dog and a great Dane. The left subclavian artery and left aortic arch remnant complex originated at the junction between the right distal aortic arch and the descending aorta and coursed dorsal to the oesophagus in a cranial direction. The attachment of the ligamentum arteriosum to the aberrant left subclavian artery was approximately 5 cm cranial to the point of origin of the aberrant left subclavian artery and left aortic arch remnant complex from the descending aorta in both dogs. This anomaly observed in both dogs is similar to an anomaly reported in humans, in which a persistent right aortic arch is found in conjunction with an aberrant left subclavian artery and a left aortic arch remnant (Kommerell's diverticulum). Surgical ligation and division of the left ligamentum arteriosum in both dogs, along with division of the left subclavian artery in the great Dane, resulted in resolution of clinical signs in both of the dogs in this report.     

Mitogen-activated protein kinases partially regulate endothelin-1-induced contractions through a myosin light chain phosphorylation-independent pathway

Endothelin (ET), derived from the endothelium of blood vessels, is a potent vasoactive peptide. Although it has been reported to be involved in cardiovascular diseases, such as hypertension, the mechanism by which ET evokes vasoconstriction is still unclear. On the other hand, p42/p44 mitogen-activated protein kinase (MAPK) and p38 MAPK are activated by a variety of growth factors and cellular stresses, respectively. However, the role of p42/p44 MAPK and p38 MAPK on the ET-1-induced vasoconstriction is not fully understood. This study was undertaken to determine whether p42/p44 MAPK and p38 MAPK participate in the regulation of vascular smooth muscle contraction by ET-1. The isometric vasoconstriction and intracellular Ca(2+) ([Ca(2+)](i)) were simultaneously measured using CAF-100. Phosphorylation of myosin light chain (MLC) and p42/p44 MAPK, p38 MAPK were determined by Western blots. In rat thoracic aorta, ET-1 induced a sustained contraction. In contrast, [Ca(2+)](i) was decreased with time. Both PD98059, an inhibitor of p42/p44 MAPK, and SB203580, an inhibitor of p38 MAPK, partially attenuated ET-1-induced contractions in concentration-dependent manners. ET-1 increased phosphorylation of both p42/p44 MAPK and p38 MAPK, and PD98059 and SB203580 completely decreased phosphorylation of p42/p44 MAPK and p38 MAPK in response to ET-1 stimulation, respectively. On the other hand, PD98059 and SB203580 did not affect MLC phosphorylation in response to ET-1 stimulation. These results indicate that p38 MAPK, as well as p42/p44 MAPK, may partially regulate the ET-1-induced contraction through a MLC phosphorylation-independent pathway.