713-4 Inhibition of Vascular Superoxide Production in Hypercholesterolemic Rabbit Aorta by L-Arginine Contributes to Restored Endothelium-dependent Relaxation

Chronic oral administration of L-arginine (L-ARG) has been shown to enhance endothelial function in cholesterol (CHOL)-fed rabbits and to reduce atherogenesis. We investigated whether modulation of endogenous NO production (as assessed by urinary NO3-excretion) by L-ARG and the inhibitor of NO synthesis, L-NAME, affects vascular superoxide (O2-) production in hypercholesterolemic rabbits. Phorbol-myristate-acetate (PMA)-stimulated O2-production from isolated aortic rings was increased in rabbits given CHOL (+159±28%) or CHOL + L-NAME (+149±37%) as compared to controls (-22±7%), and endothelium-dependent relaxations by acetylcholine were diminished in both groups. In aortic rings from rabbits given CHOL + L-ARG, PMA-induced O2-production was restored to control levels (+14±17%; p<0.05), and endothelium-dependent cholinergic relaxations were also partly restored. Urinary NO3-excretion decreased in all animals fed a CHOL-enriched diet (p<0.01). As NO inactivated by O2-is also oxidized to NO3-, this indicates a decreased endothelial production of NO. NO3-excretion was further decreased by L-NAME (p<0.05 vs. CHOL), and partly restored by L-ARG (p<0.05). We conclude that both a decreased production of NO and an enhanced breakdown of NO by O2-contribute to the diminished biological activity of endothelial NO in hypercholesterolemia. L-ARG restores endothelial function by enhancing NO formation and by protecting NO from early breakdown by O2-

Restoring vascular nitric oxide formation by l-arginine improves the symptoms of intermittent claudication in patients with peripheral arterial occlusive disease

AbstractBackground. Administration of l-arginine improves nitric oxide (NO) formation and endothelium-dependent vasodilation in atherosclerotic patients.Objectives. We investigated in this double-blind, controlled study whether prolonged intermittent infusion therapy with l-arginine improves the clinical symptoms of patients with intermittent claudication, as compared with the endothelium-independent vasodilator prostaglandin E1, and control patients.Methods. Thirty-nine patients with intermittent claudication were randomly assigned to receive 2 × 8 g l-arginine/day, or 2 × 40 μg prostaglandin E1(PGE1)/day or no hemodynamically active treatment, for 3 weeks. The pain-free and absolute walking distances were assessed on a walking treadmill at 3 km/h, 12% slope, and NO-mediated, flow-induced vasodilation of the femoral artery was assessed by ultrasonography at baseline, at 1, 2 and 3 weeks of therapy and 6 weeks after the end of treatment. Urinary nitrate and cyclic guanosine-3′, 5′-monophosphate (GMP) were assessed as indices of endogenous NO production.Results. l-Arginine improved the pain-free walking distance by 230 ± 63% and the absolute walking distance by 155 ± 48% (each p < 0.05). Prostaglandin E1improved both parameters by 209 ± 63% and 144 ± 28%, respectively (each p < 0.05), whereas control patients experienced no significant change. l-Arginine therapy also improved endothelium-dependent vasodilation in the femoral artery, whereas PGE1had no such effect. There was a significant linear correlation between the l-arginine/asymmetric dimethylarginine (ADMA) ratio and the pain-free walking distance at baseline (r = 0.359, p < 0.03). l-Arginine treatment elevated the plasma l-arginine/ADMA ratio and increased urinary nitrate and cyclic GMP excretion rates, indicating normalized endogenous NO formation. Prostaglandin E1therapy had no significant effect on any of these parameters. Symptom scores assessed on a visual analog scale increased from 3.51 ± 0.18 to 8.3 ± 0.4 (l-arginine) and 7.0 ± 0.5 (PGE1; each p < 0.05), but did not significantly change in the control group (4.3 ± 0.4).Conclusions. Restoring NO formation and endothelium-dependent vasodilation by l-arginine improves the clinical symptoms of intermittent claudication in patients with peripheral arterial occlusive disease

An endogenous inhibitor of nitric oxide synthase regulates endothelial adhesiveness for monocytes

AbstractOBJECTIVESWe sought to determine whether asymmetric dimethylarginine (ADMA) inhibits nitric oxide (NO) elaboration in cultured human endothelial cells and whether this is associated with the activation of oxidant-sensitive signaling mediating endothelial adhesiveness for monocytes.BACKGROUNDEndothelial NO elaboration is impaired in hypercholesterolemia and atherosclerosis, which may be due to elevated concentrations of ADMA, an endogenous inhibitor of NO synthase.METHODSHuman umbilical vein endothelial cells (ECV 304) and human monocytoid cells (THP-1) were studied in a functional binding assay. Nitric oxide and superoxide anion (O2−) were measured by chemiluminescence; ADMA by high pressure liquid chromatography; monocyte chemotactic protein-1 (MCP-1) by ELISA and NF-κB by electromobility gel shift assay.RESULTSIncubation of endothelial cells with ADMA (0.1 μM to 100 μM) inhibited NO formation, which was reversed by coincubation with L-arginine (1 mM). The biologically inactive stereoisomer symmetric dimethylarginine did not inhibit NO release. Asymmetric dimethylarginine (10 μM) or native low-density lipoprotein cholesterol (100 mg/dL) increased endothelial O2− to the same degree. Asymmetric dimethylarginine also stimulated MCP-1 formation by endothelial cells. This effect was paralleled by activation of the redox-sensitive transcription factor NF-κB. Preincubation of endothelial cells with ADMA increased the adhesiveness of endothelial cells for THP-1 cells in a concentration-dependent manner. Asymmetric dimethylarginine-induced monocyte binding was diminished by L-arginine or by a neutralizing anti-MCP-1 antibody.CONCLUSIONSWe concluded that the endogenous NO synthase inhibitor ADMA is synthesized in human endothelial cells. Asymmetric dimethylarginine increases endothelial oxidative stress and potentiates monocyte binding. Asymmetric dimethylarginine may be an endogenous proatherogenic molecule

Indoleamine-2,3-dioxygenase activity in experimental human endotoxemia

Background: Excessive tryptophan metabolism to kynurenine by the rate-limiting enzyme endothelial indoleamine 2,3-dioxygenase 1 (IDO) controls arterial vessel relaxation and causes hypotension in murine endotoxemia. However, its relevance in human endotoxemia has not been investigated so far. We thus aimed to study changes in blood pressure in parallel with tryptophan and kynurenine levels during experimental endotoxemia in humans. Findings: Six healthy male volunteers were given E. coli lipopolysaccharide (LPS; 4 ng/kg) as a 1-min intravenous infusion. They had levels of soluble E-Selectin and soluble vascular cell adhesion molecule-1 as well as IDO activity assessed as the kynurenine-to-tryptophan plasma ratio by liquid chromatography-tandem mass spectrometry at various time points during a 24 h time course. During endotoxemia, IDO activity significantly increased, reaching peak levels at 8 h after LPS infusion (44.0 ± 15.2 vs. 29.4 ± 6.8 at baseline, P<0.0001). IDO activity correlated inversely with the development of hypotension as shown by random effects linear regression models. Finally, IDO activity exhibited a kinetic profile similar to that of soluble endothelial-specific adhesion molecules. Conclusions: LPS is a triggering factor for the induction of IDO in men. Our findings strongly support the concept that the induction of IDO in the vascular endothelium contributes to hypotension in human sepsis

Relationship of asymmetric dimethylarginine to dialysis treatment and atherosclerotic disease

Relationship of asymmetric dimethylarginine to dialysis treatment and atherosclerotic disease. Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelial nitric oxide (NO) synthase. Its concentration is elevated in patients with end-stage renal disease (ESRD), in part because it is excreated via the kidneys. In addition, ADMA is degraded by the enzyme dimethylarginine dimethylaminohydrolase (DDAH), which hydrolyzes ADMA to L-citrulline and dimethylamine. Activity of DDAH is decreased by oxidized low density lipoprotein (LDL) or tumor necrosis factor-α (TNF-α) in vitro yielding increased levels of ADMA. Furthermore, plasma levels of ADMA are elevated in hyperhomocyst(e) inemia and in hypertensive patients on a high salt diet. Data from several experimental studies suggest that ADMA concentrations in a pathophysiologically high range (3 to 10 μmol/L) significantly inhibit vascular NO formation by NO synthase in the presence of L-arginine in isolated human blood vessels, cultured macrophages, and in cultured endothelial cells. It has been well demonstrated that ADMA accumulates in chronic renal failure. Although there is controversy concerning the absolute concentration of ADMA, all authors found a two-to sixfold increase in ADMA levels in patients in chronic renal failure as compared to controls. Different dialysis treatment strategies differentially affect ADMA levels. The presence of atherosclerosis is associated with higher ADMA levels in patients with normal renal function as well as in dialysis patients, but this phenomenon may be unrelated to renal handling of ADMA. Reduced NO elaboration secondary to accumulation of ADMA may be an important pathogenic factor for atherosclerosis in chronic renal failure and ADMA may be a new uremic toxin. Clinical studies on the effect of ADMA are needed to further elucidate its pathophysiological role in atherosclerosis and uremia