The anabolic action of intermittent parathyroid hormone on cortical bone depends partly on its ability to induce nitric oxide-mediated vasorelaxation in BALB/c mice

There is strong evidence that vasodilatory nitric oxide (NO) donors have anabolic effects on bone in humans. Parathyroid hormone (PTH), the only osteoanabolic drug currently approved, is also a vasodilator. We investigated whether the NO synthase inhibitor L-NAME might alter the effect of PTH on bone by blocking its vasodilatory effect. BALB/c mice received 28 daily injections of PTH[1-34] (80 µg/kg/day) or L-NAME (30 mg/kg/day), alone or in combination. Hindlimb blood perfusion was measured by laser Doppler imaging. Bone architecture, turnover and mechanical properties in the femur were analysed respectively by micro-CT, histomorphometry and three-point bending. PTH increased hindlimb blood flow by >30% within 10 min of injection (P < 0.001). Co-treatment with L-NAME blocked the action of PTH on blood flow, whereas L-NAME alone had no effect. PTH treatment increased femoral cortical bone volume and formation rate by 20% and 110%, respectively (P < 0.001). PTH had no effect on trabecular bone volume in the femoral metaphysis although trabecular thickness and number were increased and decreased by 25%, respectively. Co-treatment with L-NAME restricted the PTH-stimulated increase in cortical bone formation but had no clear-cut effects in trabecular bone. Co-treatment with L-NAME did not affect the mechanical strength in femurs induced by iPTH. These results suggest that NO-mediated vasorelaxation plays partly a role in the anabolic action of PTH on cortical bone

Nitric oxide synthase inhibition attenuates cardiac response to hemodilution with viscogenic plasma expander.

Background and objectivesIncreased vascular wall shear stress by elevated plasma viscosity significantly enhances the endothelial nitric oxide synthase (eNOS) activity during an acute isovolemic hemodilution. Also the modulation of plasma viscosity has effects on the cardiac function that were revealed if a left ventricular (LV) pressure-volume (PV) measurement was used. The aim of this study was to assess cardiac function responses to nitric oxide synthase (NOS) inhibitors with the presence of an elevated plasma viscosity but a low hematocrit level. Furthermore, systemic parameters were monitored in a murine model.Materials and methodsAs test group five anesthetized hamsters were administered with N(G)-nitro-L-arginine methyl ester (L-NAME), NOS inhibitor, whereas five other hamsters were used as control group without L-NAME infusion. The dosage of L-NAME was 10 mg/kg. An isovolemic hemodilution was performed by 40% of estimated blood volume with 6% w/v dextran 2000 kDa, high viscosity plasma expanders (PEs) with viscosity 6.34 cP. LV function was measured and assessed using a 1.4 Fr PV conductance catheter.ResultsThe study results demonstrated that NOS inhibition prevented the normal cardiac adaptive response after hemodilution. The endsystolic pressure increased 14% after L-NAME infusion and maintained higher than at the baseline after hemodilution, whereas it gradually decreased in the animals without L-NAME infusion. The admission of L-NAME significantly decreased the maximum rate of ventricular pressure rise (+dP/dtmax), stroke volume and cardiac output after hemodilution if compared to the control group (p&lt;0.05).ConclusionThis finding supports the presumption that nitric oxide induced by an increased plasma viscosity with the use of a high viscosity PE plays a major role in the cardiac function during an acute isovolemic hemodilution

Nitric oxide modulation of glutamatergic, baroreflex, and cardiopulmonary transmission in the nucleus of the solitary tract

The neuromodulatory effect of NO on glutamatergic transmission has been studied in several brain areas. Our previous single-cell studies suggested that NO facilitates glutamatergic transmission in the nucleus of the solitary tract (NTS). in this study, we examined the effect of the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) on glutamatergic and reflex transmission in the NTS. We measured mean arterial pressure (MAP), heart rate (HR), and renal sympathetic nerve activity (RSNA) from Inactin-anesthetized Sprague-Dawley rats. Bilateral microinjections of L-NAME (10 nmol/100 nl) into the NTS did not cause significant changes in basal MAP, HR, or RSNA. Unilateral microinjection of (RS)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA, 1 pmol/100 nl) into the NTS decreased MAP and RSNA. Fifteen minutes after L-NAME microinjections, AMPA-evoked cardiovascular changes were significantly reduced. N-methyl-D-aspartate (NMDA, 0.5 pmol/100 nl) microinjection into the NTS decreased MAP, HR, and RSNA. NMDA-evoked falls in MAP, HR, and RSNA were significantly reduced 30 min after L-NAME. To examine baroreceptor and cardiopulmonary reflex function, L-NAME was microinjected at multiple sites within the rostro-caudal extent of the NTS. Baroreflex function was tested with phenylephrine (PE, 25 mug iv) before and after L-NAME. Five minutes after L-NAME the decrease in RSNA caused by PE was significantly reduced. To examine cardiopulmonary reflex function, phenylbiguanide (PBG, 8 mug/kg) was injected into the right atrium. PBG-evoked hypotension, bradycardia, and RSNA reduction were significantly attenuated 5 min after L-NAME. Our results indicate that inhibition of NOS within the NTS attenuates baro- and cardiopulmonary reflexes, suggesting that NO plays a physiologically significant neuromodulatory role in cardiovascular regulation.Univ Texas, Hlth Sci Ctr, Dept Pharmacol, San Antonio, TX 78229 USAUniversidade Federal de São Paulo, Escola Paulista Med, Dept Physiol, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Physiol, São Paulo, BrazilWeb of Scienc

Effect of L-NAME, an inhibitor of nitric oxide synthesis, on cardiopulmonary function in human septic shock

STUDY OBJECTIVES: We tested the effects of continuous infusion of N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthesis, on cardiovascular performance and pulmonary gas exchange in patients with hyperdynamic septic shock. DESIGN: Prospective clinical study. SETTING: ICU of a university hospital. PATIENTS: Eleven critically ill patients with severe refractory septic shock. INTERVENTIONS: Standard hemodynamic measurements were made and blood samples taken before, during, and after 12 h of continuous infusion of 1 mg/kg/h of L-NAME. MEASUREMENTS AND RESULTS: Continuous infusion of L-NAME increased mean arterial pressure (MAP) from 65+/-3 (SEM) to 93+/-4 mm Hg and systemic vascular resistance (SVR) from 962+/-121 to 1,563+/-173 dyne x s x cm(-5)/m2. Parallel to this, cardiac index (CI) decreased from 4.8+/-0.4 to 3.9+/-0.4 L/min/m2 and myocardial stroke volume (SV) was reduced from 43+/-3 to 34+/-3 mL/m2. Left ventricular stroke work was increased in the first hour of L-NAME infusion from 31+/-3 to 43+/-4 g x m/m2 (all p<0.01 compared with baseline). Heart rate, cardiac filling pressures, and right ventricular stroke work did not change significantly (p>0.05). L-NAME increased the ratio of arterial PO2 to the fraction of inspired O2 from 167+/-23 to 212+/-27 mm Hg (p<0.05). Venous admixture (QVA/QT) was reduced from 19.4+/-2.6% to 14.2+/-2.1% (p<0.05) and oxygen extraction ratio increased from 21.1+/-2.4% to 25.3+/-2.7% (p<0.05). Oxygen delivery (DO2) was reduced following L-NAME, whereas oxygen uptake and arterial lactate and pH were unchanged. CONCLUSIONS: Prolonged inhibition of NO synthesis with L-NAME can restore MAP and SVR in patients with severe septic shock. Myocardial SV and CI decrease, probably as a result of increased afterload, since heart rate and stroke work were not reduced. L-NAME can improve pulmonary gas exchange with a concomitant reduction in QVA/QT. L-NAME did not promote anaerobe metabolism despite a reduction in DO2

L-NAME treatment in pigs

Abstract only availableNω-nitro-L-arginine-methyl-ester (L-NAME) inhibits the enzyme nitric oxide synthase (NOS) which generates the physiologic messenger gas, nitric oxide (NO). In addition to its role as a vasodilator NO inhibits inflammation and vascular smooth muscle cell proliferation. It was shown recently that rats fed L-NAME contain evidence of inflammation and increased collagen in their coronary vasculature when compared with control rats. We hypothesized that L-NAME treatment will cause inflammation and an increase in collagen in the coronary vasculature of pigs compared with control pigs. To test this hypothesis we have four pigs, two received L-NAME in their drinking water and two did not. We have samples of the left ventricle (LV), right ventricle (RV), and left anterior descending (LAD), left circumflex (LCX), and right (RCA) coronary arteries of the heart. Similar to studies in the rat, we will stain these samples for monocyte chemoattractant protein-1 (MCP-1), a marker of inflammation; alpha smooth muscle actin (αSMA), a marker of vascular smooth muscle, and picrosirius red (PSR) a marker of collagen. We will photograph sections of the coronary vasculature stained with these markers and use a computer image analysis system to count the amount of MCP-1, vascular smooth muscle, and collagen. Preliminary results suggest that we have insufficient statistical power to show differences in the parameters measured and need to examined greater numbers of animals.Louis Stokes Missouri Alliance for Minority Participatio

Role of nitric oxide on the electrophysiological properties of isolated rabbit atrioventricular node by extracellular field potential during atrial fibrillation

Introduction: The aim of the present study was to determine direct effects of NO modulation on protective electrophysiological properties of atrioventricular node (AV node) in the experimental model of AF in rabbit. Methods: Isolated perfused rabbit AV nodal preparations were used in two groups. In the first group (N=7), LNAME (50μM) was applied. In the second group (N=12), different concentrations of L - argenine (250 μM - 5000 μM) were added to the solution. Programmed stimulation protocols were used to quantify AV nodal conduction time, refractoriness and zone of concealment. AF protocol was executed by software with coupling intervals (ranging from 75–125 msec). Results: L-NAME had depressive effects on basic AV nodal properties. L-Arginine (250μM) had direct inhibitory effects on nodal conduction time, Wenckebach and refractoriness. Significant increases in the number of concealed beats were induced by L-Arginine (500 μM). Number of concealed beats were increased from 700.7±33.7 to 763±21 msec (P<0.05). Trend of zone of concealment prolongation in a frequency-dependent model was abrogated by Larginine (250, 5000 μM). Conclusion: NO at low concentration (in the presence of L-NAME) had facilitatory role on AV nodal properties, but at high concentration (in the presence of L-arginine) enhanced protective role of AV node during AF. Biphasic modulatory role of NO may affect protective behavior of AV node during AF. © 2011, Iranian Society of Physiology and Pharmacology. All rights reserved

Effects of NO synthase inhibitors on the synovial microcirculation in the mouse knee joint

Production of nitric oxide by the inducible NO synthase (iNOS) is known to be enhanced in chronic joint inflammation and osteoarthritis as well as aseptic loosening of joint prostheses. Initial studies yielded promising results after inhibition of the nitric oxide synthase (NOS). However, the effect of NOS inhibition has not been studied at the site of the primary function of NO, the microcirculation of the synovium in vivo. Using our recently developed model for the in vivo study of synovial microcirculation in the mouse knee joint, the effects of selective versus nonselective inhibition of iNOS were investigated by means of intravital fluorescence microscopy. After resection of the patella tendon, the synovial fatty tissue was exposed for intravital microscopy. Diameter of arterioles, functional capillary density (FCD), diameter of venules, venular red blood cell velocity and leukocyte-endothelial cell interaction were quantitatively analyzed before, and 10 and 60 min after intravenous injection of NOS inhibitors {[}selective iNOS inhibitor N-iminoethyl-L-lysine (L-NIL), and nonselective NOS inhibitor N-G-nitro-L-arginine methyl ester (L-NAME)]. Our results demonstrate that L-NAME causes a significant decrease in the arteriolar diameter and FCD associated with an increase in the leukocyte accumulation in the synovium in vivo. In contrast, L-NIL neither altered the microhemodynamics nor the leukocyte-endothelial cell interaction in the synovium, indicating its potential use for selective inhibition of iNOS in joint inflammation. Using our method, further studies will provide new insights into the unknown effect of NOS inhibition on the synovial microvasculature in inflammatory joint disease in vivo. Copyright (C) 1999 S. Karger AG, Basel

Remote preconditioning by aortic constriction: affords cardioprotection as classical or other remote ischemic preconditioning? Role of iNOS

Dose remote preconditioning by aortic constriction (RPAC) affords cardioprotection similar to classical or other remote ischemic preconditioning stimulus? Moreover study was also designed to investigate role of inducible nitric oxide synthase in remote preconditioning by aortic constriction. There are sufficient evidences that &#x22;ischemic preconditioning&#x22; has surgical applications and afford clinically relevant cardioprotection. Transient occlusion of circumflex artery, renal artery, limb artery or mesenteric artery preconditions the myocardium against ischemia reperfusion injury in case of ischemic heart disease leading to myocardial infraction. Here abdominal aorta was selected to produce RPAC. Four episodes of Ischemia-reperfusion of 5 min each to abdominal aorta produced RPAC by assessment of infract size, LDH and CK. These studies suggest RPAC produced acute (FWOP) and delayed (SWOP) cardioprotective effect. RPAC demonstrated a significant decrease in Ischemia-reperfusion induced release of LDH, CK and extent of myocardial infract size. L-NAME (10 mg/Kg i.v.), Aminoguanidine (150 mg/Kg s.c.), Aminoguanidine (300 mg/Kg s.c.), S-methyl isothiourea (3 mg/Kg i.v.), 1400W (1 mg/Kg i.v.) administered 10 min. before global ischemia reperfusion produced no marked effect. Aminoguanidine (150 mg/Kg s.c.), Aminoguanidine (300 mg/Kg s.c.), S-methyl isothiourea (3 mg/Kg i.v.), 1400W (1 mg/Kg i.v.) pretreatment after RPAC produced no significant effect on acute RPAC induced decrease in LDH, CK and infract size, whereas L-NAME (10 mg/Kg i.v.) increased RPAC induced decrease in LDH, CK and infract size. Most interesting observation is in delayed RPAC, where all NOS inhibitors pretreatment attenuate RPAC induced decrease in LDH, CK and infract size. In conclusions, &#x22;Remote preconditioning by aortic constriction&#x22; (RPAC) affords cardioprotection similar to classical or other remote ischemic preconditioning stimulus. Moreover, late or delayed phase of RPAC has been mediated by inducible nitric oxide synthase (iNOS) whereas it has not involved in acute RPAC

Effect of nitric oxide modulation on the basic and rate-dependent electrophysiological properties of AV-node in the isolated heart of rabbit: The role of adrenergic and cholinergic receptors

Introduction: Recent studies showed that nitrergic system have specific modulatory effects on electrophysiological properties of atrioventricular (AV) node. The aim of this study was to determine the effects of nitric oxide (NO) on the electrophysiological properties of isolated rabbit AV node and to investigate the role of adrenergic and cholinergic receptors in the mechanism of its action. Methods: In our laboratory, an experimental model of isolated double-perfused AV-node of rabbits weighing 1.5-2 kg was used. Specific experimental protocols of recovery, Facilitation, Fatigue and Wenckbach were applied in both control and in the presence of the drug. A total number of 35 rabbits were divided randomly into the following groups (n=7): 1) L-Arg (NO donor) (250, 750 and 1000 μmol), 2) L- NAME, a NO synthesis inhibitor (25, 50 and 100 μmol), 3) L-Arg + L- NAME, 4) Nadolol (1 μmol), 5) Atropine (3 μmol). All data were shown as mean ± SE. The level of statistical significance was set at p<0.05. Results: Our results revealed the depressant effect of L-Arg on the basic and rate-dependent electrophysiological properties of AV-node. L- NAME did not deteriorate the effects of L-Arg on the basic and rate-dependent properties, nevertheless, at high concentration (100 μmol) it had a direct inhibitory effect on the AV-node. Nadolol and atropine could prevent the effects of NO on the basic nodal characteristics and the fatigue phenomenon, respectively. Conclusion: Nitergic system can affect basic and rate-dependent electrophysiological properties of the AV-node through adrenergic and cholinergic receptors