Endogenous Nitric Oxide Synthase Research Articles

Role of Asymmetrical Dimethylarginine in Diabetic Microvascular Complications.

Microvascular complications are the leading causes of acquired blindness, end-stage renal failure, and varieties of neuropathy associated with diabetes. Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, is involved in endothelial dysfunction, oxidative stress, and inflammation associated with the progression of diabetic microvascular complications. Elevated ADMA has been detected in experimental animals and patients with diabetic microangiopathy like retinopathy, nephropathy, and neuropathy. In the review, we focus on the role of ADMA in the pathobiology of major microvascular complications of diabetes.

Arginines Plasma Concentration and Oxidative Stress in Mild to Moderate COPD.

BackgroundElevated plasma concentrations of the endogenous nitric oxide synthase (NOS) inhibitor asymmetric dimethylarginine (ADMA) have been observed in respiratory conditions such as asthma and cystic fibrosis. Since oxidative stress has been shown to increase the activity of arginine methylating enzymes, hence increased ADMA synthesis, and to reduce ADMA degrading enzymes, hence increased ADMA concentrations, we assessed methylated arginines concentrations in chronic obstructive pulmonary disease (COPD), a disease characterized by increased oxidative stress.MethodsPlasma arginine, ADMA and symmetric dimethylarginine (SDMA), oxidative stress markers (thiobarbituric acid reactive substances, TBARS, and plasma proteins SH, PSH) and antioxidants (taurine and paraoxonase 1, PON1, activity) were measured in 43 COPD patients with mild (n = 29) or moderate (n = 14) disease and 43 age- and sex-matched controls.ResultsTBARS significantly increased with COPD presence and severity (median 2.93 vs 3.18 vs 3.64 μmol/L, respectively in controls, mild and moderate group, p<0.0001 by ANOVA) whereas PSH decreased (6.69±1.15 vs 6.04±0.85 vs 5.33±0.96 μmol/gr prot, p<0.0001 by ANOVA). Increased ADMA/arginine ratio, primarily due to reduced arginine concentrations, was also observed with COPD presence and severity (median 0.0067 vs 0.0075 vs 0.0100, p<0.0001 by ANOVA). In multiple logistic regression analysis, only TBARS (OR 0.44, 95% CI 0.25–0.77; p = 0.0045) and ADMA/Arginine ratio (OR 1.72, 95% CI 2.27–13.05; p = 0.02) were independently associated with COPD severity.ConclusionCOPD presence and severity are associated with increased oxidative stress and alterations in arginine metabolism. The reduced arginine concentrations in COPD may offer a new target for therapeutic interventions increasing arginine availability.

Upregulation of CBS/H2S system contributes to asymmetric dimethylarginine-triggered protection against the neurotoxicity of glutamate to PC12 cells by inhibiting NOS/NO pathway

Glutamate-induced neurotoxicity involves in overproduction of nitric oxide (NO) and oxidative stress. Our previous data demonstrated that asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase (NOS) inhibitor, has a protective effect against glutamate-induced neurotoxicity. Hydrogen sulfide (H2S), the third endogenous gaseous mediator, has potential therapeutic value for oxidative stress-induced neural damage. Therefore, we hypothesized that ADMA provides protection against the neurotoxicity of glutamate by regulating endogenous H2S generation. In the present study, we found that ADMA prevented glutamate-triggered decrease in endogenous H2S generation in PC12 cells and reversed glutamate-induced suppression in the expression and activity of cystathionine-β-synthetase (CBS), the predominant enzymatic source of H2S in PC12 cells. Furthermore, AOAA, a potent inhibitor of CBS, significantly abolished the protective action of ADMA against glutamate-induced neurotoxicity to PC12 cells. We also showed that ADMA suppressed glutamate-elicited NOS excessive activation and NO overproduction in PC12 cells. These data indicate that the protection of ADMA against glutamate-induced neurotoxicity is by promoting endogenous H2S generation, resulting from suppression in NOS excessive activation and NO overproduction. These findings provide a novel mechanism underlying the protection of ADMA against glutamate-induced neurotoxicity.

Association of Asymmetric Dimethylarginine With Acute Pancreatitis-Induced Hyperglycemia.

The objective of this study was to investigate the relationship between asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, oxidative-nitrosative damage, and glucoregulation in acute pancreatitis (AP). The study evaluated serum levels of ADMA, nitrotyrosine, and urinary 8-hydroxydeoxyguanosine in 40 male patients hospitalized for AP at baseline and at 2 and 10 days of treatment, respectively. The patients were classified into a mild and a moderately severe AP group (MAP and MSAP, respectively) according to Atlanta classification criteria. Glycemic status was evaluated by a 75-g oral glucose tolerance test 1 month after AP onset. Forty age-matched healthy subjects served as control subjects. Significant decrease of ADMA and increased levels of nitrotyrosine and urinary 8-hydroxydeoxyguanosine were found in MSAP, but not in MAP at baseline, with ADMA correction toward control levels at the 10th day of treatment. Fructosamine was found to significantly influence ADMA levels (r = -0.362, P = 0.002). After AP recovery, either impaired glucose tolerance or diabetes was identified with the oral glucose tolerance test in 10.5% and 92.8% of patients with MAP and MSAP, respectively. Insufficient inhibition of nitric oxide synthesis, through reduced bioavailability of ADMA, might be a novel significant contributory factor to the severity of AP and subsequent development of hyperglycemia.

BAY 41-2272 Treatment Improves Acetylcholine-Induced Aortic Relaxation in L-NAME Hypertensive Rats

Hypertension, an emerging problem of recent era, and many pathophysiological factors are participating to produce the disease. Nitric oxide (NO) is an important constituent to ameliorate hypertensive condition. Inhibition of endogenous NO synthase by L-NG-Nitroarginine methyl ester (L-NAME) was responsible for generating hypertension in rats. BAY 41-2272 (5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]-pyrimidin-4-ylamine), a soluble guanylyl cyclase activator, restricts rise of blood pressure and shows cardioprotective activity. The aim of the present study was to analyze effect of short-term BAY 41-2272 treatment on blood pressure and vascular function. Male Wistar rats were randomly divided into three groups such as control (group-A), hypertensive (group-B), and BAY 41-2272-treated hypertensive (group-C) rats. Normal saline was administered intramuscularly to control rats for last 3 days (days 40, 41, and 42) of total 42 days treatment, whereas rats of group-B and group-C were treated with L-NAME hydrochloride in drinking water at 50 mg/kg body weight daily for 42 days. Also, normal saline and BAY 41-2272 were administered for last 3 days at two different dosages at 1 and 3 mg/kg body weight/day intramuscularly to group-B and group-C rats, respectively. Administration of BAY 41-2272 for 3 days was not sufficient enough to decrease mean arterial pressure of hypertensive rats significantly. BAY at both the treatment dosages significantly ameliorate acetylcholine-induced maximal aortic relaxation compared with BAY-untreated hypertensive rats. Findings of the present study indicate that even shorter period of BAY 41-2272 treatment (3 days) improves vascular relaxation.

Role of asymmetric dimethylarginine in rheumatological disease's cardiovascular complications

Inflammation that is an independent predictor of cardiovascular disease plays a important role in rheumatological diseases. Asymmetric dimethylarginine (ADMA), an inhibitor of endogenous nitric oxide synthase, contributes to the inflammatory syndrome. Elevated ADMA levels in many rheumatological diseases suggest that ADMA might play an important role in the rheumatological diseases and cardiovascular complication. In this review, we discuss the role of ADMA in rheumatological disease's cardiovascular complications. Key words: Arginine/ME; Rheumatic diseases/CO/ME; Immune system diseases/CO/ME; Review

DDAH1 plays dual roles in PM2.5 induced cell death in A549 cells

BackgroundDimethylarginine dimethylaminohydrolase 1 (DDAH1) is an enzyme that can degrade asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase (NOS) inhibitor. Emerging evidence suggests that alterations in the ADMA–DDAH1 pathway are involved in environmental pollution induced airway inflammation. However, the role of DDAH1 in protection against cytotoxicity of ambient airborne particulate matter is unclear. MethodsWe examined the influence of DDAH1 expression on oxidative stress and cell apoptosis in human type II alveolar epithelial A549 cells exposed to PM2.5 (particulate matter with an aerodynamic diameter less than 2.5μM). ResultsWe found that PM2.5 exposure for 48h significantly decreased DDAH1 expression. However, knockdown of DDAH1 prior to PM2.5 exposure actually attenuated the cytotoxicity of PM2.5. Cytoprotection in DDAH1 deficient cells was due to increased reactive oxygen species, activation of PI3K–AKT and mitogen-activated protein kinase (MAPK) pathways, subsequent activation of nuclear factor erythroid-2-related factor 2 (Nrf2) and this caused a subsequent reduction in PM2.5 induced oxidative stress relative to control. DDAH1 depletion also repressed the induction of inducible NOS (iNOS) in PM2.5-exposed cells and knockdown of iNOS protected cells against PM2.5 induced cell death. Interestingly, overexpression of DDAH1 also exerted a protective effect against the cytotoxicity of PM2.5 and this was associated with a reduction in oxidative stress and upregulation of the anti-apoptotic protein Bcl-2. ConclusionsOur data indicate that DDAH1 plays dual roles in protection against cytotoxicity of PM2.5 exposure, apparently by limiting PM2.5 induced oxidative stress. General significanceOur findings reveal new insights into the role(s) of the DDAH1/ADMA in pulmonary protection against airborne pollutants. This article is part of a Special Issue entitled Air Pollution, edited by Wenjun Ding, Andrew J. Ghio and Weidong Wu.

IL-4 promotes asymmetric dimethylarginine accumulation, oxo-nitrative stress, and hypoxic response–induced mitochondrial loss in airway epithelial cells

Obesity is known to increase asthma risk and severity. Increased levels of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, are associated with mitochondrial toxicity, asthma, and metabolic syndrome. IL-4 upregulates the expression of protein arginine methyltransferases, which are essential for ADMA formation. Importantly, cross-talk between IL-4, ADMA, and mitochondrial dysfunction could explain how obesity and IL-4 can synergize to exacerbate allergic inflammation. We sought to investigate how IL-4, a key asthma-associated cytokine, can influence ADMA-related effects on lungs. BEAS2B (bronchial epithelial) cells were treated with IL-4 followed by ADMA and investigated for oxo-nitrative stress and resultant mitochondrial toxicity after 48hours by using flow cytometry, confocal imaging, immunoblotting, and fluorimetric assays. IL-4-induced mitotoxicity in BEAS2B cells was significantly higher in the presence of exogenous ADMA. IL-4 treatment led to proteolytic degradation of dimethylarginine dimethylaminohydrolase 2, which catabolizes ADMA. IL-4 pretreatment was associated with increased intracellular ADMA accumulation and increased ADMA-induced mitotoxicity. Airway epithelial cells treated with IL-4 followed by ADMA showed exaggerated oxo-nitrative stress and potent induction of the cellular hypoxic response, despite normoxic conditions. The hypoxic response was associated with reduced mitochondrial function but was reversible by overexpression of the mitochondrial biogenesis factor, mitochondrial transcription factor A. We conclude that IL-4 promotes intracellular ADMA accumulation, leading to mitochondrial loss through oxo-nitrative stress and hypoxic response. This provides a novel understanding of how obesity, with high ADMA levels, and asthma, with high IL-4 levels, might potentiate each other and highlights the potential of mitochondrial-targeted therapeutics in obese subjects with asthma.

Plasma concentrations of three methylated arginines, endogenous nitric oxide synthase inhibitors, in schizophrenic patients undergoing antipsychotic drug treatment

Plasma concentration of three methylated arginines, endogenous nitric oxide synthase inhibitors, is not studied in schizophrenic patients. The purpose of this study was to determine plasma concentrations of NG-monomethyl-l-arginine (l-NMMA), NG,NG-dimethyl-l-arginine (ADMA), NG,NG’-dimethyl-l-arginine (SDMA), and l-arginine in 56 male and 45 female schizophrenic patients undergoing antipsychotic drug treatment versus those of 39 male and 24 female healthy controls. Plasma concentrations of methylated arginines and l-arginine were measured using newly developed high performance liquid chromatography with fluorescence detection which we previously reported. Methylated arginine levels were slightly but significantly higher in schizophrenic patients. l-Arginine levels and the l-arginine/(ADMA+l-NMMA) ratio were higher in schizophrenic patients than in healthy controls. It is considered that pharmacological treatment of schizophrenic patients may lower methylated arginine levels that are increased by the disease, and increase l-arginine levels, eliciting an improvement in nitric oxide (NO) bioavailability.

Peroxynitrite Produced via Nitric Oxide Synthesis in Isolated Cardiac Mitochondria

It is controversial whether or not isolated mitochondria contain an endogenous nitric oxide synthase (NOS). NOS catalyzes the production of the gaseous signaling molecule NO•(nitric oxide). During ischemia/reperfusion (IR) injury, NO•reacts with superoxide (O2•−) to form the oxidant peroxynitrite (ONOO−), which can cause tissue damage and initiate deleterious post-translational modifications of cellular proteins. The aim of this project is to detect the production of ONOO-in isolated mitochondria under conditions of oxidative/nitrosative stress, and to determine whether the ONOO-production is dependent on the presence of endogenous mitochondrial NOS. We hypothesize that isolated mitochondria contain endogenous NOS. Mitochondrial suspensions were incubated at pH 7.15, and treated with succinate, a complex II substrate, followed by menadione, a redox cycler that has been shown to generate large amounts of O2•−. In physiological buffer, L-tyrosine is oxidized by ONOO−to form the fluorescent dimer dityrosine (diTyr). We found that exposing isolated mitochondria to CaCl2, followed by succinate resulted in an increase in diTyr fluorescence signal over a 30 second period, indicating that the simulated IR injury condition increases mitochondrial ONOO− production. To verify the NOS-mediated production of ONOO−, mitochondria were incubated with the NOS inhibitors L-NAME/L-NNA, a superoxide dismutase (SOD) mimetic, TEMPOL, and PTIO, a potent global NO•scavenger. L-NAME and L-NNA both abolished the succinate and menadione-induced increase in ONOO−. The global NO• scavenger PTIO also abolished the production of ONOO-. TEMPOL partially attenuated the ONOO- levels by converting O2•− to H2O2. These results suggest that mitochondrial NOS is responsible for the production of ONOO-during simulated oxidative/nitrosative stress.

Nebivolol ameliorates asymmetric dimethylarginine-induced vascular response in rat aorta via β3 adrenoceptor-mediated mechanism

Background: Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase (NOS) inhibitor, induces endothelial dysfunction. Nebivolol, a highly selective β1-adrenergic receptor (AR) blocker, is the only beta-blocker known to induce vascular production of nitric oxide. Objective: The present study was designed to evaluate the effect and mechanism of nebivolol on ADMA-induced vascular response in rat aorta in vitro. Methods: In vitro, the effects of nebivolol and ADMA on resting tone or contraction induced by phenylephrine (PE, 10−6 mol/L) and relaxation induced by acetylcholine (Ach, 10−10–10−5 mol/L) were evaluated. Results: ADMA in a concentration-dependent manner increased the resting and PE-induced tone and reduced Ach-induced relaxation. Nebivolol inhibited the ADMA-induced enhancements in tone and reversed the effects of ADMA on Ach-induced relaxation. These effects of nebivolol were blocked by selective β3 receptor blocker cyanopindolol (1 μM), but not by selective β2 receptor blocker butoxamine (50 μM). Conclusions: Nebivolol ameliorates the ADMA-induced vascular responses in rat aorta, at least in part, by mechanisms involving β3 adrenoceptor.

DDAH1 deficiency promotes intracellular oxidative stress and cell apoptosis via a miR-21-dependent pathway in mouse embryonic fibroblasts

Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase (NOS) inhibitor, is degraded by dimethylarginine dimethylaminohydrolase 1 (DDAH1). Emerging evidence suggests that plasma ADMA accumulation, DDAH1 activity/expression reduction, and microRNA-21 (miR-21) upregulation are linked to disease pathology, but the mechanisms remain largely unknown. In the present study, we assessed the potential role of the ADMA–DDAH1–miR-21 pathway in the regulation of the cellular redox state and apoptosis using wild-type (WT) and DDAH1-knockout (KO) immortalized mouse embryonic fibroblasts (MEFs). DDAH1 deficiency significantly increased ADMA levels, enhanced cellular oxidative stress, and rendered cells more vulnerable to apoptosis induced by tert-butyl hydroperoxide (tBHP) or A23187. However, treatment with exogenous ADMA (1–80μM) for 24h or for a prolonged period (10μM, 10 passages) in WT MEFs had no marked effect on intracellular reactive oxygen species (ROS) and apoptosis sensitivity. Interestingly, miR-21 expression was significantly increased, by 4 fold, in DDAH1−/− MEFs, and the induction of miR-21 by DDAH1 deficiency was dependent on oxidative stress and NF-κB activation. Inhibition of DDAH1 activity by PD 404182 also increased miR-21 expression. Furthermore, inhibition of miR-21 with a lentiviral vector in DDAH1−/− MEFs significantly upregulated SOD2 expression and the attenuated oxidative stress and apoptosis induced by tBHP or A23187. Taken together, our results suggest that DDAH1 not only acts as an enzyme degrading ADMA but also controls cellular oxidative stress and apoptosis via a miR-21-dependent pathway.

Nitric oxide synthase-dependent immune response against gram negative bacteria in a crustacean, Litopenaeus vannamei.

Nitric oxide (NO) is a short-lived radical generated by nitric oxide synthases (NOS). NO is involved in a variety of functions in invertebrates, including host defense. In previous studies, we isolated and sequenced for the first time the NOS gene from hemocytes of Panulirus argus, demonstrating the inducibility of this enzyme by lipopolysaccharide in vitro e in vivo. Hyperimmune serum was obtained from rabbits immunized with a P. argus -NOS fragment of 31 kDa produced in Escherichia coli, which specifically detected the recombinant polypeptide and the endogenous NOS from lobster hemocytes by western blotting and immunofluorescence. In the present work, we demonstrate that the hyperimmune serum obtained against P. argus NOS also recognizes Litopenaeus vannamei NOS in hemocytes by western blotting and immunofluorescence. Our data also show that while the hemolymph of L. vannamei has a strong antibacterial activity against the Gram negative bacteria Aeromonas hydrophila, the administration of the anti NOS serum reduce the natural bacterial clearance. These results strongly suggest that NOS is required for the shrimp immune defense toward Gram negative bacteria. Therefore, the monitoring of induction of NOS could be an important tool for testing immunity in shrimp farming.

Does the ADMA/DDAH/NO pathway modulate early regression of left ventricular hypertrophy with esmolol?

Hypertensive left ventricular hypertrophy (LVH) is a maladaptive response to chronic pressure overload and a strong independent risk factor for cardiovascular disease. Regression of LVH is associated with improved prognosis. Regression of LVH with antihypertensive therapy (angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, beta-blockers, calcium channel blockers, and diuretics) has been reported, although only after long-term treatment. Asymmetrical dimethylarginine (ADMA), the most potent endogenous NO synthase inhibitor, is emerging as an important cardiovascular risk factor in patients with arterial hypertension and LVH, and dimethylarginine dimethylaminohydrolase (DDAH) is the mechanism that most frequently leads to accumulation of ADMA (plasma ADMA is cleared in small part by renal excretion, although the bulk of ADMA is degraded by DDAH). Left ventricular mass is strongly modulated by the NO system. As an important inhibitor of the bioavailability of NO, ADMA is an underlying mechanism of LVH. Beta-blockers can induce regression of LVH and reduced plasma ADMA levels. Oxidative stress is increased in patients with LVH, and this in turn increases generation of ADMA. In a previous preclinical study of spontaneously hypertensive rats, we found that short-term treatment (48h) with esmolol reverses early LVH, increases the bioavailability of NO, and improves antioxidant status in plasma. Therefore, we propose that the ADMA/DDAH/NO pathway could modulate early regression of LVH with esmolol.

Asymmetric dimethylarginine and homocysteine in exudative age-related macular degeneration

Introduction: The oxidative stress has been proposed as an important case of exudative age-related macular degeneration (E-ARMD). The aim of the present study was to investigate homocysteine (Hcy), asymmetric dimethylarginine (ADMA) and oxidized low-density lipoprotein cholesterol (Ox-LDL-C) levels, the factors involved in oxidative stress, in the patients with E-ARMD. Methods: In a cross-sectional study, 45 patients with E-ARMD were compared with 45 sex- and age-matched healthy controls. The levels of biochemical factors, Hcy, ADMA, and of Ox-LDL were estimated by standard methods in both study groups. Results: The levels of Hcy (15.4 ± 7.2 vs. 10.7 ± 3.7 μM, P = 0.001), Ox-LDL (52.2 ± 13.8 vs. 37.8 ± 10.8 U/l, P = 0.001), and ADMA (0.84 ± 0.23 vs. 0.71 ± 0.26 μM, P = 0.012) were significantly higher in the patients with E-ARMD than those in the controls. In the patient group, there was a positive and significant correlation between serum Ox-LDL and Hcy concentrations (r = 0.719, P = 0.001), but no correlation was found between serum ADMA and Ox-LDL (r = 0.010, P = 0.900) and also between serum Hcy and ADMA levels (r = -0.070, P = 0.600). Conclusion: The high levels of Hcy as an oxidant agent and ADMA as an endogenous nitric oxide synthase inhibitor can lead to increase Ox-LDL levels, and they may have important roles in oxidative stress, which can be a trigger in E-ARMD.

Low urinary citrulline/arginine ratio associated with blood pressure abnormalities and arterial stiffness in childhood chronic kidney disease

Arginine (ARG) and citrulline (CIT) are essential for nitric oxide (NO) synthesis. Their metabolites are interrelated, and involved in blood pressure (BP) control, chronic kidney disease (CKD), and cardiovascular disease (CVD). Although CVD is the leading cause of mortality in CKD, little is known about subclinical CVD in early-stage childhood CKD. Twenty-four–hour ambulatory BP monitoring and arterial stiffness assessment allows the earlier possible detection of subclinical CVD. We investigated whether urinary CIT and ARG metabolites and their ratios are correlated with BP load and vascular abnormalities in children and adolescents with early-stage CKD. We enrolled 55 pediatric patients with mild-to-moderate CKD. Seventy percent (30/43) had at least one out of BP load abnormality on ambulatory BP monitoring, mainly increased asleep systolic BP (SBP) load (40%), asleep SBP or diastolic BP load > 95th percentile (40%), and nocturnal SBP nondipping (35%). Low urinary CIT level and CIT/ARG ratio were associated with BP load abnormalities in children with early CKD. Urinary CIT/ARG ratio was correlated with arterial stiffness, represented as pulse-wave velocity and augmentation index. SBP and diastolic BP loads were negatively correlated with urinary CIT, ARG, asymmetric dimethylarginine (an endogenous NO synthase inhibitor), and CIT/ARG ratio, while positively associated with dimethylamine/asymmetric dimethylarginine ratio and pulse-wave velocity. Early assessments of BP load abnormalities, urinary biomarkers in the CIT-ARG-NO pathway, and arterial stiffness parameters should increase early preventive care toward decreasing hypertension and CV remodeling in pediatric CKD.

The role of adipose tissue asymmetric dimethylarginine/dimethylarginine dimethylaminohydrolase pathway in adipose tissue phenotype and metabolic abnormalities in subtotally nephrectomized rats.

The lipodystrophy-like phenotype has been suggested in early chronic kidney disease (CKD). It includes adipose tissue atrophy, systemic insulin resistance (IR), dyslipidemia and ectopic lipid accumulation. To elucidate its pathogenesis, we investigated the role of two uremic toxins that affect insulin sensitivity: an endogenous nitric oxide synthase inhibitor, and asymmetric dimethylarginine (ADMA) and indoxyl sulfate (IS). Six-week-old Sprague-Dawley rats were rendered CKD by subtotal nephrectomy (Nx) and compared with sham-operated rats. Cultured 3T3-L1 fibroblasts were differentiated into mature adipocytes with or without ADMA. Transgenic (Tg) mice overexpressing each isoform of ADMA degrading enzyme, dimethylarginine dimethylaminohydrolase 1 (DDAH1) and DDAH2 were subject to Nx and their phenotypes were investigated. In Nx rats, IR was evident and insulin stimulation failed to activate insulin signaling in adipose tissues. Adipose tissue weight, adipocyte size and adipocyte differentiation marker expressions decreased as a consequence of IR in Nx. Tissue lipid content in the liver and muscle increased in Nx rats. Tissue levels of ADMA, IS and oxidative stress increased in the adipose tissue of Nx rats. Both DDAH1 and DDAH2 expressions decreased, and a putative IS receptor, aryl hydrocarbon receptor, expression increased in the adipose tissue of Nx rats. ADMA inhibited adipocyte differentiation, triglyceride accumulation and insulin signaling, which were reversed by pretreatment with cGMP. In each type of Tg mice overexpressing DDAH1 or DDAH2, all lipodystrophy-like phenotypes induced by Nx were reversed. In mild CKD, dysregulation of the ADMA/DDAH pathway in adipose tissue triggers lipodystrophy-like phenotype changes, including ectopic fat depositions.

Plasmodium Infection Is Associated with Impaired Hepatic Dimethylarginine Dimethylaminohydrolase Activity and Disruption of Nitric Oxide Synthase Inhibitor/Substrate Homeostasis.

Inhibition of nitric oxide (NO) signaling may contribute to pathological activation of the vascular endothelium during severe malaria infection. Dimethylarginine dimethylaminohydrolase (DDAH) regulates endothelial NO synthesis by maintaining homeostasis between asymmetric dimethylarginine (ADMA), an endogenous NO synthase (NOS) inhibitor, and arginine, the NOS substrate. We carried out a community-based case-control study of Gambian children to determine whether ADMA and arginine homeostasis is disrupted during severe or uncomplicated malaria infections. Circulating plasma levels of ADMA and arginine were determined at initial presentation and 28 days later. Plasma ADMA/arginine ratios were elevated in children with acute severe malaria compared to 28-day follow-up values and compared to children with uncomplicated malaria or healthy children (p<0.0001 for each comparison). To test the hypothesis that DDAH1 is inactivated during Plasmodium infection, we examined DDAH1 in a mouse model of severe malaria. Plasmodium berghei ANKA infection inactivated hepatic DDAH1 via a post-transcriptional mechanism as evidenced by stable mRNA transcript number, decreased DDAH1 protein concentration, decreased enzyme activity, elevated tissue ADMA, elevated ADMA/arginine ratio in plasma, and decreased whole blood nitrite concentration. Loss of hepatic DDAH1 activity and disruption of ADMA/arginine homeostasis may contribute to severe malaria pathogenesis by inhibiting NO synthesis.

Effect of rosiglitazone on asymmetric dimethylarginine metabolism in thioacetamide-induced acute liver injury

Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, is metabolized in the liver by dimethylarginine dimethylaminohydrolase (DDAH). We aimed to investigate the effect of rosiglitazone, a peroxysome proliferator-activated receptor-gamma (PPAR-γ) agonist, on ADMA metabolism in acute liver injury. Male Sprague Dawley rats were injected thioacetamide (TAA; 500mgkg−1) intraperitoneally in order to induce acute liver injury. ADMA, SDMA and arginine levels were determined in plasma by the HPLC. Liver DDAH activity and malondialdehyde (MDA) levels were measured by spectrophotometric procedures. TAA injection caused marked increases in ALT and AST activities. Plasma ADMA levels were increased, while arginine levels and arginine/ADMA ratio were decreased. Liver DDAH activity was significantly diminished and MDA levels were elevated. In another group of animals which were treated with a PPAR-γ agonist (rosiglitazone, 5mgkg−1) daily via gastric intubation for a week prior to TAA injection, significant recoveries in DDAH activity and antioxidant status were observed when compared with solely TAA-injected animals. Rosiglitazone pretreatment improved the plasma arginine/ADMA ratio.Our findings indicated that PPAR-γ agonist rosiglitazone beneficially influenced hepatic metabolism of ADMA in TAA-induced acute liver damage.

Levels of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, and risk of coronary artery disease: A meta-analysis based on 4713 participants.

Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of endothelial nitric oxide synthase by competing with L-arginine. As a result, the expression of nitric oxide decreases and endothelial dysfunction occurs. Studies have evaluated the association between the serum ADMA level and risk of coronary artery disease. However, conflicting results have been obtained. Pubmed, Web of Science, Embase, Ovid, Cochrane databases were searched to identify eligible studies published in English until December 2014. Association was assessed on the basis of weighted mean differences (WMD) with 95% confidence intervals (CIs). Publication bias was analysed using Begg's and Egger's tests. Sensitivity analysis was performed to evaluate result stability. A total of 16 case-control studies with 2939 patients and 1774 controls were included in the meta-analysis. Pooled result indicated that patients with coronary artery disease yielded a higher ADMA level than healthy controls (WMD: 0.248, 95% CI: 0.156-0.340; p = 1.16 e-7). Sensitivity analysis suggested that our meta-analysis result was stable. Subgroup analysis found a similar pattern in patients with myocardial infarction (WMD: 0.397, 95% CI: 0.112-0.683; p = 0.0106), stable angina pectoris (WMD: 0.197, 95% CI: 0.031-0.364; p = 0.02) and unstable angina pectoris (WMD: 0.857, 95% CI: 0.293-1.420; p = 0.003). Meta-analysis results indicated that an increased ADMA level is associated with an increased risk of coronary artery disease.