Vasoconstrictor Serotonin Research Articles

Functional sympatholysis in mouse skeletal muscle involves sarcoplasmic reticulum swelling in arterial smooth muscle cells.

The vasoconstrictive effect of sympathetic activity is attenuated in contracting skeletal muscle (functional sympatholysis), allowing increased blood supply to the working muscle but the underlying mechanisms are incompletely understood. The purpose of this study was to examine α‐adrenergic receptor responsiveness in isolated artery segments from non‐exercised and exercised mice, using wire myography. Isometric tension recordings performed on femoral artery segments from exercised mice showed decreased α‐adrenergic receptor responsiveness compared to non‐exercised mice (logEC50 −5.2 ± 0.04 M vs. −5.7 ± 0.08 M, respectively). In contrast, mesenteric artery segments from exercised mice displayed similar α‐adrenergic receptor responses compared to non‐exercised mice. Responses to the vasoconstrictor serotonin (5‐HT) and vasodilator isoprenaline, were similar in femoral artery segments from non‐exercised and exercised mice. To study sarcoplasmic reticulum (SR) function, we examined arterial contractions induced by caffeine, which depletes SR Ca2+ and thapsigargin, which inhibits SR Ca2+‐ATPase (SERCA) and SR Ca2+ uptake. Arterial contractions to both caffeine and thapsigargin were increased in femoral artery segment from exercised compared to non‐exercised mice. Furthermore, 3D electron microscopy imaging of the arterial wall showed SR volume/length ratio increased 157% in smooth muscle cells of the femoral artery from the exercised mice, whereas there was no difference in SR volume/length ratio in mesenteric artery segments. These results show that in arteries surrounding exercising muscle, the α‐adrenergic receptor constrictions are blunted, which can be attributed to swollen smooth muscle cell SR’s, likely due to increased Ca2+ content that is possibly reducing free intracellular Ca2+ available for contraction. Overall, this study uncovers a previously unknown mechanism underlying functional sympatholysis.

Heartwood extract of Rhus verniciflua Stokes and its active constituent fisetin attenuate vasoconstriction through calcium-dependent mechanism in rat aorta.

Rhus verniciflua Stokes (RVS) exert cardiovascular protective activity by promoting blood circulation, but its active ingredients and underlying mechanism have yet to be identified. This study investigated the vascular effects of RVS, focusing on vasoconstriction and smooth muscle Ca(2+) signaling. RVS heartwood extract attenuated contraction of aortic rings induced by the vasoconstrictors serotonin and phenylephrine, and inhibited the Ca(2+) signaling evoked by serotonin in vascular smooth muscle cells. Subsequent activity-guided fractionation identified fisetin as an active constituent exerting a Ca(2+) inhibitory effect. Fisetin could inhibit major Ca(2+) mobilization pathways including extracellular Ca(2+) influx mediated by the L-type voltage-gated Ca(2+) channel, Ca(2+) release from the intracellular store and store-operated Ca(2+) entry. In accordance with Ca(2+) inhibitory effect, fisetin attenuated vasoconstriction by serotonin and phenylephrine. These results suggest that the anticontractile effect, which is presumably mediated by inhibition of Ca(2+) signaling, may contribute to the improvement of blood circulation by RVS.

Increased plasma serotonin metabolite 5-hydroxyindole acetic acid concentrations are associated with impaired systolic and late diastolic forward flows during cardiac cycle and elevated resistive index at popliteal artery and renal insufficiency in type 2 diabetic patients with microalbuminuria.

Although lower extremity arterial disease is frequently accompanied by diabetes mellitus, the association of circulating biomarkers with flow components during the cardiac cycle in lower-leg arteries has yet to be fully elucidated. We enrolled 165 type 2 diabetic patients with normal ankle-brachial index (ABI 1.0-1.4), comprising 106 normoalbuminuric and 59 microalbuminuric patients, and 40 age-matched nondiabetic subjects consecutively admitted to our hospital. Serum high sensitivity C-reactive protein (hsCRP) level and plasma von Willebrand factor ristocetin cofactor activity (VWF) and vasoconstrictor serotonin metabolite 5-hydroxyindole acetic acid (5-HIAA) concentrations were measured. An automatic device was used to measure ABI and brachial-ankle pulse wave velocity (baPWV). Flow components during the cardiac cycle, total flow volume, and resistive index at popliteal artery were evaluated using gated magnetic resonance imaging. Although estimated glomerular filtration rate (eGFR), early diastolic flow reversal, heart rate, and ABI were similar between the groups, diabetic patients had higher log hsCRP (p<0.001), VWF (p<0.001), 5-HIAA (p=0.002), resistive index (p<0.001) and baPWV (p<0.001) and lower systolic (p=0.026) and late diastolic (p<0.001) forward flows and total flow volume (p<0.001) than nondiabetic subjects. Multivariate analyses demonstrated that 5-HIAA in microalbuminuric patients showed higher associations with systolic and late diastolic forward flows during the cardiac cycle, total flow volume and resistive index at popliteal artery, and eGFR compared to normoalbuminuric patients. In microalbuminuric patients, 5-HIAA was a significant independent determinant among these factors. Thus, increased plasma 5-HIAA levels are involved in the pathogenesis of impaired blood flow in lower extremities and renal insufficiency in diabetic patients with microalbuminuria.

Associations of plasma von Willebrand factor ristocetin cofactor activity and 5-hydroxyindole acetic acid concentrations with blood flow in lower-leg arteries in Japanese type 2 diabetic patients with normal ankle-brachial index

AimsTo evaluate the associations of circulating levels of proinflammatory molecules and endothelial factors with blood flow in lower-leg arteries in diabetic patients with normal ankle-brachial index (ABI>0.9). MethodsWe enrolled 123 type 2 diabetic patients with normal ABI and 30 age-matched nondiabetic subjects consecutively admitted to our hospital. Flow volume and resistive index, an index of peripheral vascular resistance, at the popliteal artery were evaluated using gated two-dimensional cine-mode phase-contrast magnetic resonance imaging. An automatic device was used to measure ABI and brachial-ankle pulse-wave velocity (baPWV) for evaluation of arterial stiffness. Plasma soluble intercellular adhesion molecule-1 (sICAM-1) and monocyte chemoattractant protein-1 (MCP-1) concentrations, serum high-sensitivity C-reactive protein (hsCRP) levels, plasma von Willebrand factor ristocetin cofactor activity (VWF), and plasma vasoconstrictor serotonin metabolite 5-hydroxyindole acetic acid (5-HIAA) concentrations were measured. ResultsDiabetic patients had higher baPWV (P<.0001), resistive index (P<.0001), sICAM-1 (P<.0001), MCP-1 (P=.0224), log hsCRP (P<.0001), VWF (P<.0001), 5-HIAA (P=.0015), and lower blood flow (P<.0001) than nondiabetic subjects. VWF (P=.0019) or 5-HIAA (P=.0011), but not sICAM-1, MCP-1, and log hsCRP, was negatively correlated with blood flow in diabetic patients. A multivariate analysis revealed that the significant independent determinants of blood flow were hypertension, use of renin–angiotensin system inhibitors, VWF and 5-HIAA (r2=0.198, P<.0001) in diabetic patients. ConclusionsPlasma VWF and 5-HIAA concentrations are associated with blood flow and are involved in the pathogenesis of impaired peripheral circulation due to higher arterial stiffness and greater vascular resistance in lower-leg arteries in diabetic patients with normal ABI.

Manipulation of vascular function by blood flukes?

Schistosomes (blood flukes) are long lived, intravascular parasites that afflict ~ 200 million people worldwide. Here we review the potential ability of these parasites to exert control on local vascular physiology. We examine schistosome kallikrein-like proteins that drive vasodilation. We review biogenic amine metabolism in the parasites that involve the vasodilator histamine and its receptors and the vasoconstrictor serotonin and its receptor. Schistosomes can trigger the release of histamine from host cells and can import serotonin. We consider the ability of schistosomes to generate and release the eicosanoid vasodilators PGD 2 and PGE 2 and the vasoconstrictors LTB 4 and LTC 4. The literature on nitric oxide metabolism in these blood flukes is assessed. Finally the potential impact of other schistosome metabolic processes (e.g. exogenous adenosine generation and acetylcholine degradation) on vascular function is appraised. An increased understanding of these processes could lead to novel anti-parasitics as well as new therapies to treat vascular dysfunction.

Deletion of the serotonin transporter in rats disturbs serotonin homeostasis without impairing liver regeneration

The serotonin transporter is implicated in the uptake of the vasoconstrictor serotonin from the circulation into the platelets, where 95% of all blood serotonin is stored and released in response to vascular injury. In vivo studies indicated that platelet-derived serotonin mediates liver regeneration after partial hepatectomy. We have recently generated serotonin transporter knockout rats and demonstrated that their platelets were almost completely depleted of serotonin. Here we show that these rats exhibit impaired hemostasis and contain about 1-6% of wild-type serotonin levels in the blood. Despite the marked reduction of serotonin levels in blood and platelets, efficient liver regeneration and collagen-induced platelet aggregation occur in rats lacking the serotonin transporter. These results provide evidence that liver regeneration is not dependent on the release of serotonin from platelets. Our findings indicate that very low levels of serotonin in blood are sufficient for liver regeneration.

Inversion of the response to serotonin in rats with traumatic shock

Normally serotonin reduced blood pressure. It was shown that in rats with traumatic shock its hypotensive effect was transformed into hypertensive one. In vitro serotonin exhibited a slight vasoconstrictor effect on isolated rat aorta, while 24 h after injury, the strength of aortic contractions in response to serotonin increased 2.2 times. Desensitization of glucocorticoid receptors caused by injection of high doses of dexamethasone (3 mg/kg) to rats for 5 days led to similar changes in serotonin effect. We hypothesized that inversion of the response to serotonin in shock was caused by increased activity and/or expression of vasoconstrictor serotonin receptors in blood vessels.

Medical marvels The 100 greatest advances in medicine

Eugene Straus, professor emeritus of medicine at the State University of New York Health Science Center, has followed up his interesting biography, Rosalyn Yalow, Nobel laureate: her life and work in medicine (1), with Medical marvels: the 100 greatest advances in medicine. Coauthored with journalist Alex Straus, with interesting and stimulating black-and-white illustrations by Bette Korman, the volume examines the science and art of many key medical advances throughout history, selected based on the criteria that they changed the course of medicine, relieved suffering, furthered our understanding of health and disease, and culminated in improvements in disease prevention, diagnosis, or therapeusis. Each of the 100 advances is described concisely within individual 3- to 5-page chapters, thus serving as a launching pad for the reader to acquire additional information on specific topics of interest. Naturally, the breakthroughs that most scientists, physicians, and lay readers will be familiar with appear. These include the discovery of viruses, oncogenes, antibiotics, and stem cells; smallpox vaccination; gene therapy; and, of course Watson, Crick, Wilkins, and Franklin’s identification of the structure of DNA and the elucidation of the human genetic code, regarded as the most important biological experiments in history. The authors also examine the evolution of some of the experimental and diagnostic approaches that researchers and physicians have come to rely on daily, including: history taking and the physical exam, surgical anesthesia, tissue culture, the x-ray, the ECG, and PCR. The authors revisit Antonie van Leeuwenhoek’s construction of the single-lens microscope and William Beaumont’s examination via a perforation in the anterior wall of the stomach of a patient following a shotgun wound, which provided the earliest insights into the digestive process and its regulation in vivo. This led to the work of Ivan Pavlov, whose experiments with dogs provided great insight into the mechanisms of salivation, pancreatic function, hunger, and satiation and identified the phenomenon now known as classical conditioning. As an additional resource, the authors include as part of their discussion of solid organ transplantation contact information for national organizations that assist patients in need of transplants. In the 1940s, as a naval medical corpsman, I had the great fortune to be among the first to administer a new drug — penicillin — to casualties from the Pacific Island invasions. At that time the results obtained from using penicillin were astounding and never before equaled by any therapeutic agent for infections. Alexander Fleming and Howard Florey’s work toward the discovery and commercial production of penicillin rightly appears on Straus’s list, and this drug represents a true medical marvel. Throughout the text each chapter is admixed with others concerned with social and economic problems associated with socialized medicine, medical ethics, health maintenance organizations, patient advocacy, and related subjects. These include the evolution of the doctor-patient relationship into one of a provider and customer, the management of addictive behavior, and the underrepresentation of minorities and women in the pursuit of medicine. As interesting and important as the sections on the social aspects of medicine are, they distract from the flow of scientific information and would have better constituted a separate section of the book. This is not to diminish the importance of these sections, but rather to note that they interrupt the sequence of summaries of medical research–related accomplishments. Based on my own experience in medicine and research since 1953, I would have included several additional advances in the physical sciences, akin to some of those highlighted by John Gribbin in The scientists: a history of science told through the lives of its greatest inventors (2), along with the elucidation of the structure of the psychotropic serum vasoconstrictor serotonin and the research of Erwin Chargaff that actually laid the groundwork for the discovery of the double-helix structure of DNA. I would hope to see these featured if a second edition of this volume is released. Thus, we have an interesting, richly illustrated book for both the layman and scientist who wish to develop broad baseline information on major medical advances. However, more than simply a compendium of breakthroughs in medical technology, Medical marvels intertwines the social and ethical aspects of the practice of medicine with the science, impressing upon the reader the dedication of individuals committed to enhancing the quality of human life. The text is strongly recommended as interesting, informative, and stimulating reading.

Evaluation of the Serotonin Receptor Blockers Ketanserin and Methiothepin on the Pulmonary Hypertensive Responses of Broilers to Intravenously Infused Serotonin

The pathogenesis of pulmonary hypertension remains incompletely understood. Many factors have been implicated; however, there has been great interest in the potent pulmonary vasoconstrictor serotonin (5-HT) due to episodes of primary pulmonary hypertension in humans triggered by serotoninergic appetite-suppressant drugs. Pulmonary hypertensive patients have elevated blood 5-HT levels and pulmonary vasoconstriction induced by 5-HT is believed to be mediated through 5-HT1B/1D and 5-HT2A receptors that are expressed by pulmonary smooth muscle cells. The vascular remodeling associated with pulmonary hypertension also appears to require the serotonin transporter. We investigated the roles of 5-HT receptor blockers on the development of pulmonary hypertension induced by infusing 5-HT i.v. in broilers. For this purpose, we treated broilers with the selective 5-HT2A receptor antagonist ketanserin (5 mg/ kg of BW) or with the nonselective 5-HT1/2 receptor antagonist methiothepin (3 mg/kg of BW). Receptor blockade was followed by infusion of 5-HT while recording pulmonary arterial pressure and pulmonary arterial blood flow. The results demonstrate that methiothepin, but not ketanserin, eliminated the 5-HT-induced pulmonary hypertensive responses in broilers. The 5-HT2A receptor does not, therefore, appear to play a role in the 5-HT-induced pulmonary hypertensive responses in broilers. Methiothepin did not inhibit pulmonary vascular contractility per se, because the pulmonary hypertensive response to the thromboxane A2 mimetic U44069 remained intact in methiothepin-treated broilers. Methiothepin will be a useful tool for evaluating the role of 5-HT in the pathogenesis of pulmonary hypertension syndrome (ascites) as well as the onset of pulmonary hypertension triggered by inflammatory stimuli such as bacterial lipolysaccharide.

ATP-sensitive K+ currents in cerebral arterial smooth muscle: pharmacological and hormonal modulation.

Calcitonin gene-related peptide (CGRP), hypoxia, and synthetic activators of ATP-sensitive potassium (KATP) channels (e.g., pinacidil and levcromakalim) cause dilation of cerebral arteries that are attenuated by the KATP channel inhibitor glibenclamide. We have identified and characterized KATP currents in smooth muscle cells isolated from rabbit cerebral arteries, using the whole cell configuration of the patch-clamp technique. Pinacidil (10 microM) and levcromakalim (10 microM) increased glibenclamide-sensitive currents about sixfold in cells dialyzed with 0.1 mM ATP. Glibenclamide-sensitive currents in the presence of pinacidil were potassium selective, voltage independent, and reduced about threefold by elevating intracellular ATP from 0.1 to 3.0 mM. External tetraethylammonium and 4-aminopyridine at millimolar concentrations reduced pinacidil-induced currents, whereas iberiotoxin, a blocker of calcium-activated potassium channels, had no effect. The vasoconstrictors serotonin and histamine also inhibited pinacidil-induced currents. The vasodilators CGRP and adenosine, in contrast, increased glibenclamide-sensitive potassium currents. We conclude that cerebral artery smooth muscle cells have KATP channels that are regulated by endogenous vasoconstrictors and vasodilators. We propose that these channels are involved in the dilation of cerebral arteries to CGRP and synthetic vasodilators.

Vasoreactivity and structure of human coronary arteries irradiated by excimer laser.

In this study the effects of excimer laser (308 nm) angioplasty on arteries have been studied. We report histology and vasoreactivity properties of human atherosclerotic and normal coronary arteries and bovine coronary arteries after exposure to pulsed excimer laser irradiation. Segments of isolated and pharmacologically active human coronary arteries were obtained within 5-8 hr postmortem. Segments of bovine arteries were obtained from fresh hearts. Side branches were ligated and vessels perfused with Krebs-Ringer's-bicarbonate solution in a perfusion apparatus to maintain their viability. Excimer laser irradiation was performed using a fused silica fiber advanced through the artery within a polyethylene cannula to assist with steerability and to protect against perforation. The fiber delivered a total of 40-60 J at its tip at 100 mJ/pulse at 15 pulses/sec. A total of 5 human atherosclerotic, 2 human normal, and 8 bovine arteries were used. Ablation of plaque was accomplished with remarkable ease and little resistance. Histologic studies showed clear-cut wedges inscribed by the catheter along the arterial wall without charring or thermal effects of coagulation necrosis. Vasoreactivity was assessed by measuring flow changes during perfusion before and after excimer angioplasty using a transient challenge with the vasoconstrictor serotonin. All excimer-irradiated arteries showed no increase in vasoreactivity during the challenge and showed preservation of relaxation properties following the challenge. In conclusion, coronary artery angioplasty by excimer laser appears to allow for effective plaque ablation with simultaneous preservation of structure and pharmacological properties of arteries.

Serotonin-mediated acute insulin resistance in the perfused rat hindlimb but not in incubated muscle: A role for the vascular system

We have recently shown that the vasoconstrictor serotonin (5-HT) inhibits oxygen uptake in perfused hindlimb possibly due to vascular shunting. Thus in the present study the effect of 5-HT on insulin-mediated glucose uptake was assessed. Rat hindlimbs were perfused at constant flow with medium containing 8.3 mM glucose and a tracer amount of 2-deoxy-D-[1- 3H]glucose (2DG) with and without 10 μM 5-HT, 15 nM insulin and a combination of the two. 5-HT inhibited insulin-mediated stimulation of glucose uptake by 30.4% when added after insulin and 34.4% when added before insulin. In addition, 5-HT inhibited insulin-mediated 2DG uptake by perfused muscles with inhibition ranging from 32% (soleus) to 80% (extensor digitorum longus). The effects of 5-HT on insulin-mediated glucose uptake were partially reversed by vasodilation with carbachol. In contrast to the results for the hindlimb, 10 μM 5-HT had no significant effect on either basal glucose uptake or the stimulation of glucose uptake mediated by 15 nM insulin by isolated incubated soleus or extensor digitorum longus muscles. It is concluded that 5-HT impairs insulin-mediated glucose uptake in the perfused rat hindlimb that may derive from vascular shunting not apparent when muscles are incubated with 5-HT in vitro. These findings may have implications for the link between hypertension and diabetes.

Effect of serotonin on intracellular free calcium of rat cerebrovascular smooth muscle cells in culture

Smooth muscle cells were dissociated from conducting cerebral arteries of adult rats and maintained in culture for 2-4 days. The calcium-sensitive fluorescent probe, fura-2, was used to study the effect of the vasoconstrictor serotonin (5-HT) on the level of free intracellular Ca2+ in these cells. The baseline level of free intracellular calcium was 39 +/- 3.6 nM. In 74 out of 110 cells, 5-HT application transiently increased the free Ca2+ content. This effect was dose-dependent and was suppressed by nanomolar concentrations of the 5-HT2 receptor antagonist, ketanserin. The 5-HT induced rise in free intracellular calcium was not prevented by the presence of Co2+, La3+, or nifedipine, blockers of voltage-sensitive calcium channels. These results indicate that 5-HT mobilizes intracellular Ca2+ in cultured smooth muscle cells derived from the rat cerebrovasculature. The mobilization of intracellular Ca2+ appears to be triggered by a 5-HT2 type receptor, although further pharmacological experiments are required to verify this hypothesis.

Isoflurane causes endothelium-dependent inhibition of contractile responses of canine coronary arteries.

The authors sought to determine if isoflurane would attenuate effects of three different types of vasoconstrictors on isolated segments of canine epicardial coronary arteries removed from healthy dogs. As the endothelium has a major role in regulating epicardial coronary artery tone, and as it modulates the effect of many vasoactive substances, experiments were conducted both on normal rings and on rings whose endothelium had been mechanically removed. In addition, the endothelium is thought to be damaged in human atherosclerosis. Rings were suspected in organ chambers filled with modified Krebs-Ringer bicarbonate solution, aerated with 95% oxygen and 5% carbon dioxide, and connected to strain gauges for the measurement of isometric tension. Isoflurane 2.3% (1.5 MAC in the dog) was added to the aerating gas mixture in half the preparations, while the other rings served as control. The vasoconstrictors serotonin, phenylephrine, or prostaglandin F2 alpha were added in increasing concentrations to the bath solution. In the presence of endothelium, vasoconstrictor evoked contractions were attenuated by isoflurane. Maximal tension generated by prostaglandin F2 alpha in untreated rings was 114 +/- 18% (mean +/- SEM) of a reference contraction, while, following isoflurane, it was 46 +/- 8% (P less than 0.005). In the absence of endothelium, isoflurane attenuated neither prostaglandin F2 alpha nor serotonin evoked contraction, and had decreased effectiveness against phenylephrine mediated contraction (P less than 0.001). It is concluded that isoflurane attenuates vasoconstrictor-evoked contraction of isolated canine epicardial coronary arteries, and that this effect is mediated by the endothelium.