3-nitrotyrosine and thiobarbituric-acid

Chronic neuroinflammation is a common characteristic of neurodegenerative diseases, and lipopolysaccharide (LPS) signaling is linked to glutamate-nitric oxide-Na,K-ATPase isoforms pathway in central nervous system (CNS) and also causes neuroinflammation. Intermittent fasting (IF) induces adaptive responses in the brain that can suppress inflammation, but the age-related effect of IF on LPS modulatory influence on nitric oxide-Na,K-ATPase isoforms is unknown. This work compared the effects of LPS on the activity of α1,α2,3 Na,K-ATPase, nitric oxide synthase gene expression and/or activity, cyclic guanosine monophosphate, 3-nitrotyrosine-containing proteins, and levels of thiobarbituric acid-reactive substances in CNS of young and older rats submitted to the IF protocol for 30 days. LPS induced an age-related effect in neuronal nitric oxide synthase activity, cyclic guanosine monophosphate, and levels of thiobarbituric acid-reactive substances in rat hippocampus that was linked to changes in α2,3-Na,K-ATPase activity, 3-nitrotyrosine proteins, and inducible nitric oxide synthase gene expression. IF induced adaptative cellular stress-response signaling pathways reverting LPS effects in rat hippocampus of young and older rats. The results suggest that IF in both ages would reduce the risk for deficits on brain function and neurodegenerative disorders linked to inflammatory response in the CNS.

Topics: Aging; Animals; Fasting; Hippocampus; Lipopolysaccharides; Male; Neurodegenerative Diseases; Neurogenic Inflammation; Nitric Oxide; Nitric Oxide Synthase; Nucleotides, Cyclic; Oxidative Stress; Rats, Wistar; Sodium-Potassium-Exchanging ATPase; Thiobarbiturates; Tyrosine

Recent studies show that endogenous hydrogen sulfide (H(2)S) plays an anti-inflammatory role in the pathogenesis of airway inflammation. This study investigated whether exogenous H(2)S may counteract oxidative stress-mediated lung damage in allergic mice. Female BALB/c mice previously sensitized with ovalbumin (OVA) were treated with sodium hydrosulfide (NaHS) 30 min before OVA challenge. Forty eight hours after antigen-challenge, the mice were killed and leukocyte counting as well as nitrite plus nitrate concentrations were determined in the bronchoalveolar lavage fluid, and lung tissue was analysed for nitric oxide synthase (NOS) activity, iNOS expression, superoxide dismutase (SOD), catalase, glutathione reductase (GR) and glutathione peroxidase (GPx) activities, thiobarbituric acid reactive species and 3-nitrotyrosine containing proteins (3-NT). Pre-treatment of OVA-sensitized mice with NaHS resulted in significant reduction of both eosinophil and neutrophil migration to the lungs, and prevented the elevation of iNOS expression and activity observed in the lungs from the untreated allergic mice, although it did not affect 3-NT. NaHS treatment also abolished the increased lipid peroxidation present in the allergic mouse lungs and increased SOD, GPx and GR enzyme activities. These results show, for the first time, that the beneficial in vivo effects of the H(2)S-donor NaHS on allergic airway inflammation involve its inhibitory action on leukocyte recruitment and the prevention of lung damage by increasing endogenous antioxidant defenses. Thus, exogenous administration of H(2)S donors may be beneficial in reducing the deleterius impact of allergic pulmonary disease, and might represent an additional class of pharmacological agents for treatment of chronic pulmonary diseases.

Topics: Animals; Catalase; Female; Glutathione Peroxidase; Glutathione Reductase; Hydrogen Sulfide; Hypersensitivity; Leukocytes; Lung; Mice; Mice, Inbred BALB C; Nitric Oxide Synthase; Oxidative Stress; Sulfides; Superoxide Dismutase; Thiobarbiturates; Tyrosine

We have previously shown that treating streptozotocin-induced diabetic rats, an animal model of type 1 diabetes, with Ilepatril (an inhibitor of neutral endopeptidase and angiotensin converting enzyme (ACE)) improves vascular and neural functions. In this study we sought to determine the effect of Ilepatril treatment of high fat fed/low dose streptozotocin-diabetic rats, a model for type 2 diabetes, on vascular and neural complications. Following 8 weeks on a high fat diet rats were treated with 30 mg/kg streptozotocin (i.p.) and after 4 additional weeks a group of these rats was treated for 12 weeks with Ilepatril followed by analysis of neural and vascular functions. Included in these studies were age-matched control rats and rats fed a high fat diet and treated with or without Ilepatril. Diabetic and diet induced obese rats have characteristics of insulin resistance, slowing of nerve conduction velocity, thermal hypoalgesia, reduction in intraepidermal nerve fiber density in the hindpaw and impairment in vascular relaxation to acetylcholine and calcitonin gene-related peptide in epineurial arterioles of the sciatic nerve. Treatment with Ilepatril was efficacious in improving all of these endpoints although improvement of insulin resistance in diabetic rats was minimal. These studies suggest that dual inhibition of angiotensin converting enzyme and neutral endopeptidase activity of type 2 diabetic rats is an effective approach for treatment of diabetic neural and vascular complications.

Topics: Adipose Tissue; Animals; Arterioles; Blood Glucose; Blood Vessels; Body Weight; Carbohydrate Metabolism; Diabetes Complications; Diabetes Mellitus, Experimental; Diet, High-Fat; Dose-Response Relationship, Drug; Enzyme Inhibitors; Glucose Tolerance Test; Glutathione; Heterocyclic Compounds, 3-Ring; Insulin; Lens, Crystalline; Leptin; Male; Muscle, Skeletal; Nerve Fibers; Nervous System; Nociception; Organ Size; Rats; Rats, Sprague-Dawley; Sciatic Nerve; Superoxides; Thiobarbiturates; Tyrosine; Vasodilation

Brain death induces multiple-organ dysfunction, with undesirable consequences for organ transplantation. However, the mechanisms are not completely clear. In the hearts, lungs, livers, and kidneys of rats, we investigated whether brain death leads to changes in nitric oxide (NO) production or to the formation of nitrotyrosine (the footprint of peroxynitrite, formed from NO and superoxide) or to lipid peroxidation products. To produce a rat model of brain death, we inflated a subdurally placed balloon catheter. We used the Griess reaction to assay plasma nitrite and nitrate. Proteolytic digestion followed by high-performance liquid chromatography (HPLC) with electrochemical detection determined nitrotyrosine formation in the tissues. Tissues were also examined immunohistochemically with anti-nitrotyrosine antibody. We used a thiobarbituric acid method to assay lipid peroxidation. An intense, transient hemodynamic activation occurred at the onset of brain death (heart rate, 496 beats/min; mean arterial pressure (AP), 181 mm Hg; dP/dt(max), 11,500 mm Hg/sec). A constant hypotensive phase (mean AP, 50 mm Hg; dP/dt(max), 2,674 mm Hg/sec) followed. Plasma concentration of nitrite plus nitrate remained unchanged 2 hours after brain death (32.8 +/- 1.5 vs 31.3 +/- 2.2 micromol/liter at zero time). Neither HPLC nor immunohistochemistry detected significant nitrotyrosine formation in the tissues. We detected no increase in lipid peroxidation products.Our results indicate that changes in the generation of reactive nitrogen and active oxygen species do not play an important role in post-brain-death organ dysfunction, at least not at the early stage.

Topics: Animals; Biomarkers; Brain Death; Chromatography, High Pressure Liquid; Heart Transplantation; Hemodynamics; Lipid Peroxidation; Liver; Liver Transplantation; Male; Multiple Organ Failure; Myocardium; Nitrates; Nitric Oxide; Nitrites; Rats; Rats, Wistar; Thiobarbiturates; Tissue Donors; Tyrosine