Nitro-oxidative Stress Research Articles

The Role of Oxidative Stress, Mitochondrial Function, and Autophagy in Diabetic Polyneuropathy.

Diabetic polyneuropathy (DPN) is the most frequent and prevalent chronic complication of diabetes mellitus (DM). The state of persistent hyperglycemia leads to an increase in the production of cytosolic and mitochondrial reactive oxygen species (ROS) and favors deregulation of the antioxidant defenses that are capable of activating diverse metabolic pathways which trigger the presence of nitro-oxidative stress (NOS) and endoplasmic reticulum stress. Hyperglycemia provokes the appearance of micro- and macrovascular complications and favors oxidative damage to the macromolecules (lipids, carbohydrates, and proteins) with an increase in products that damage the DNA. Hyperglycemia produces mitochondrial dysfunction with deregulation between mitochondrial fission/fusion and regulatory factors. Mitochondrial fission appears early in diabetic neuropathy with the ability to facilitate mitochondrial fragmentation. Autophagy is a catabolic process induced by oxidative stress that involves the formation of vesicles by the lysosomes. Autophagy protects cells from diverse stress factors and routine deterioration. Clarification of the mechanisms involved in the appearance of complications in DM will facilitate the selection of specific therapeutic options based on the mechanisms involved in the metabolic pathways affected. Nowadays, the antioxidant agents consumed exogenously form an adjuvant therapeutic alternative in chronic degenerative metabolic diseases, such as DM.

In vitro nitro-fatty acid release from Cys-NO2-fatty acid adducts under nitro-oxidative conditions

Stress situations are characterized by a rise in reactive oxygen (ROS) and nitrogen (RNS) species levels. Nitro-fatty acids (NO2-FAs), or nitroalkenes, are produced by the interaction of RNS and unsaturated fatty acids, stored in cells, mostly as part of protein-adducted NO2-FAs, and are esterified in complex lipids. These molecules, which have been shown to play a pivotal role as anti-inflammatory and pro-survival players, have been widely characterized in animal systems. Recently, it has been reported that NO2-FAs play an important role in plant defense against several stress conditions. Furthermore, a significant increase in NO2-FA levels has been observed under various inflammatory and stressful conditions in both animal and plant systems. In this study, we describe the in vitro release of NO2-FAs from protein-adducts under nitro-oxidative stress conditions. The findings of this study highlight the ability of hydrogen peroxide and peroxynitrite, as representative ROS and RNS molecules induced under stress conditions, to oxidize cysteine-adducted NO2-FAs, which is followed by the release of free nitroalkenes. This release may be partly responsible for the increase in NO2-FA content observed under different stressful conditions in both animal and plant systems as well as the activation of antioxidant and anti-inflammatory properties attributed to these molecules.

CA3 hippocampal field: Cellular changes and its relation with blood nitro-oxidative stress reveal a balancing function of CA3 area in rats exposed to repetead restraint stress.

Rats exposed to repeated restraint stress exhibit structural and functional deficits in hippocampus that are similar to those observed in patients with depressive illnesses. Blood corticosterone concentrations are proportionally increased with catalase and glutathione-peroxidase activity and are inversely proportional with 3-nitrotyrosine concentrations.Cytochrome c oxidase, adenosin tryphosphatase and monoamine oxidase activities of CA3 hippocampal field mark a stress-time dependent decrease. Acridine-orange labeling of the CA3 field reveals an enhancing green fluorescence of glyocites in stress conditions. After three days of restraint stress, the secretory activity of CA3 neurons did not show significant decrease, and neurons appeared with normal shapes and distribution. CA3 neurons after seven days of restraint stress have marked a slight decrease of secretory activity. In contrast to a well-preserved histological appearance of the CA3 neurons, local and blood stress-related reactions are observed. CA3-glial activation and disturbance of blood oxidative homeostasis are tandem processes during three and seven days of RS. This study depicts the balancing role of CA3 area in time-varying stress conditions.

State insulin resistance, the intensity of endogenous intoxication in patients with chronic pancreatitis against obesity treatment in the dynamics of L-carnitine and L-glutathione

Aim. To study the L-carnitine and L-glutathione influence on the clinical course of chronic pancreatitis (CP) in patients with obesity. Materials and methods. Three representative groups of patients were formed to determine the effectiveness of treatment. The average age of patients was (41.7±4.2) years. The common control group included 30 practically healthy persons (PHP) of the corresponding age and gender. The control group (group 1 (K), 25 persons), the basic group 2 (group 2 (A), 30 persons), the basic group 3 (group 3 (B) 25 persons). Results. After applying the proposed therapy in CP patients (group b) in the course of the disease compared with the group K showed an improvement of health, reduction of asthenic-vegetative, intoxication, and dyspepsia syndromes signs. After 2 weeks of therapy asthenic-vegetative syndrome was absent. Analysis of indicators in 3 months after treatment indicated that patients of the 2nd and the 3rd were achieved stable remission of CP, as no patient visited doctors for medical aid again. But, in 3 months after treatment 2 patients of the K group (10%) were admitted again, 5 patients (25%) sought medical help repeatedly concerning exacerbation of CP symptoms to the gastroenterologists. The result of the complex therapy with L-glutathione and L-carnitine addition positive effect on the IR syndrome is defined in our study a significant decrease of body mass index in patients of the 2nd and the 3rd groups during a 60 days period respectively: 7.5% and 8.1% (p<0.05). Conclusions. A specific method of patients’ treatment with CP on the background of obesity, which includes basic therapy together with the using of L-glutathione and L-carnitine combination, helps to eliminate clinical and biochemical signs of chronic pancreatitis exacerbation, to reduce the degree of endogenous intoxication, oxidative and nitro-oxidative stress, to normalize steadily the glycemic and insulin level, to reduce body weight.

Biochemical and Physiological Changes in Response to Salinity in Leaves and Roots of Two Durum Wheat (Triticum durum Desf.) Genotypes

Abstract Salt stress is a major abiotic stress that limits agricultural productivity in many regions of the world. To understand the molecular basis of the salt stress response, two wheat (Triticum durum Desf.) cultivars, Karim and Azizi, which are of agronomic significance in Tunisia, were grown under non-saline and saline conditions (100 mM). Leaves and roots of control and salt-stressed plants were harvested after 11 days of salt treatment. Karim cultivar may behave as a typical Na+ include, which compartmentalizes Na+ within the leaf cell vacuoles where it could be used as an osmoticum to lower the osmotic potential necessary for the maintenance of the plant hydric status. While, accumulation of K+ was greater in Karim cultivar compared to Azizi, in both organs, presenting an important manifestation of salinity tolerance. Significant changes in metabolism of antioxidative system were observed, with an increase in protein tyrosine nitration, which indicates that salinity stress induces a nitro-oxidative stress.

Nitric oxide signaling and its crosstalk with other plant growth regulators in plant responses to abiotic stress.

Nitric oxide (NO) is a free radical molecule involved in an array of functions under physiological and adverse environmental conditions. As other free radical molecules, NO biological action depends on its cellular concentration, acting as a signal molecule when produced at low concentration or resulting in cellular damage when produced at sufficiently high levels to trigger nitro-oxidative stress. Over the last decade, significant progress has been made in characterizing NO metabolism and action mechanism, revealing that diverse biosynthetic routes can generate this free radical in plants and its action mainly occurs through posttranslational modification (nitration and S-nitrosylation) of target proteins. Intricate crosstalk networks between NO and other signaling molecules have been described involving phytohormones, other second messengers, and key transcription factors. This review will focus on our current understanding of NO interplay with phytohormones and other plant growth regulators under abiotic stress conditions.

Effect of peritoneal dialysis fluid containing osmo-metabolic agents on human endothelial cells.

BackgroundThe use of glucose as the only osmotic agent in peritoneal dialysis (PD) solutions (PDSs) is believed to exert local (peritoneal) and systemic detrimental actions, particularly in diabetic PD patients. To improve peritoneal biocompatibility, we have developed more biocompatible PDSs containing xylitol and carnitine along with significantly less amounts of glucose and have tested them in cultured Human Vein Endothelial Cells (HUVECs) obtained from the umbilical cords of healthy (C) and gestational diabetic (GD) mothers.MethodsPrimary C- and GD-HUVECs were treated for 72 hours with our PDSs (xylitol 0.7% and 1.5%, whereas carnitine and glucose were fixed at 0.02% and 0.5%, respectively) and two glucose-based PDSs (glucose 1.36% or 2.27%). We examined their effects on endothelial cell proliferation (cell count), viability (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay), intracellular nitro-oxidative stress (peroxynitrite levels), Vascular Cell Adhesion Molecule-1 and Intercellular Adhesion Molecule-1 membrane exposure (flow cytometry), and HUVEC-monocyte interactions (U937 adhesion assay).ResultsCompared to glucose-based PDSs, our in vitro studies demonstrated that the tested PDSs did not change the proliferative potential both in C- and GD-HUVECs. Moreover, our PDSs significantly improved endothelial cell viability, compared to glucose-based PDSs and basal condition. Notably, glucose-based PDSs significantly increased the intracellular peroxynitrite levels, Vascular Cell Adhesion Molecule-1 and Intercellular Adhesion Molecule-1 membrane exposure, and endothelial cell–monocyte interactions in both C- and GD-HUVECs, as compared with our experimental PDSs.ConclusionPresent results show that in control and diabetic human endothelial cell models, xylitol–carnitine-based PDSs do not cause cytotoxicity, nitro-oxidative stress, and inflammation as caused by hypertonic glucose-based PDSs. Since xylitol and carnitine are also known to favorably affect glucose homeostasis, these findings suggest that our PDSs may represent a desirable hypertonic solution even for diabetic patients in PD.

37 - Resonance Raman Studies on Manganese Porphyrin Detection and Redox State in Endothelial Cells

Manganese(III) cationic ortho N-substituted pyridylporhyrins (MnPs) act as efficient antioxidants catalyzing superoxide dismutation and accelerating peroxynitrite reduction. Importantly, MnPs reach mitochondria and protect them in different animal models of disease, furthermore, MnTnBuOE-2-PyP 5+ is presently in Phase I/II clinical trial as a normal tissue radioprotectant. An LC-MS/MS -based method for MnPs quantitation and subcellular distribution has been reported; however, a direct method to evaluate the uptake and redox state of MnPs in vivo has not been developed. Here, we applied a spectroscopic technique, based on Resonance Raman (RR) confocal microscopy, in order to observe the intracellular accumulation of two potent MnP-based lipophilic SOD mimics, MnTnBuOE-2-PyP 5+ and MnTnHex-2-PyP 5+ within endothelial cells. RR spectra confirmed that MnPs were incorporated into endothelial cells where the reduced Mn(II)P is the predominant oxidation state, confirming that the Mn(III)P added can be reduced by intracellular components, in particular those of the mitochondrial electron transport chain. Thus, the incubation of Mn(III)P with isolated mitochondria or submitochondrial particles (SMP), in the presence of succinate as substrate, revealed the reduction to Mn(II)P, which was in part affected by the inhibitors of electron transport chain. Such data support the action of MnPs as efficient redox active compounds in mitochondria. Additional studies were performed in order to evaluate the cytoprotective capacity of MnPs by exposing the endothelial cells to nitro-oxidative stress induced by the peroxynitrite donor, SIN-1. We observed a preservation of normal mitochondrial function, attenuation of cell damage and prevention of apoptotic cell death. The data presented herein introduce a novel application of the RR spectroscopy for the direct detection of MnPs and their redox state in vivo , and helps to rationalize its antioxidant capacity in biological systems.

“The roots” of selenium toxicity: A new concept

ABSTRACTElevated levels of selenium (Se) cause toxicity in non-accumulator plant species. The primary reasons for toxic Se effect have been considered to be selenoprotein accumulation and oxidative stress. However, based on our recent paper in Plant Cell Reports and previous literature data we suggest that disturbances in the homeostasis of both reactive oxygen and nitrogen species result in selenium-induced nitro-oxidative stress, contributing to toxicity. The most characteristic symptom of Se exposure is the inhibited root elongation which is partly caused by hormonal disturbances. Our recent paper suggests the involvement of cytokinin in selenium stress sensing of the root systemAltogether, the aim of this Addendum is to present reactive nitrogen species and phytohormones as new players in plant selenium toxicity.

Glucagon‐like peptide‐1 receptor signalling reduces microvascular thrombosis, nitro‐oxidative stress and platelet activation in endotoxaemic mice

Excessive inflammation in sepsis causes microvascular thrombosis and thrombocytopenia associated with organ dysfunction and high mortality. The present studies aimed to investigate whether inhibition of dipeptidyl peptidase-4 (DPP-4) and supplementation with glucagon-like peptide-1 (GLP-1) receptor agonists improved endotoxaemia-associated microvascular thrombosis via immunomodulatory effects. Endotoxaemia was induced in C57BL/6J mice by a single injection of LPS (17.5 mg kg-1 for survival and 10 mg kg-1 for all other studies). For survival studies, treatment was started 6 h after LPS injection. For all other studies, drugs were injected 48 h before LPS treatment. Mice treated with LPS alone showed severe thrombocytopenia, microvascular thrombosis in the pulmonary circulation (fluorescence imaging), increased LDH activity, endothelial dysfunction and increased markers of inflammation in aorta and whole blood (leukocyte-dependent oxidative burst, nitrosyl-iron haemoglobin, a marker of nitrosative stress, and expression of inducible NOS). Treatment with the DPP-4 inhibitor linagliptin or the GLP-1 receptor agonist liraglutide, as well as genetic deletion of DPP-4 (DPP4-/- mice) improved all these parameters. In GLP-1 receptor-deficient mice, both linagliptin and liraglutide lost their beneficial effects and improvement of prognosis. Incubation of platelets and cultured monocytes (containing GLP-1 receptor protein) with GLP-1 receptor agonists inhibited the monocytic oxidative burst and platelet activation, with a GLP-1 receptor-dependent elevation of cAMP levels and PKA activation. GLP-1 receptor activation in platelets by linagliptin and liraglutide strongly attenuated endotoxaemia-induced microvascular thrombosis and mortality by a cAMP/PKA-dependent mechanism, preventing systemic inflammation, vascular dysfunction and end organ damage. This article is part of a themed section on Redox Biology and Oxidative Stress in Health and Disease. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v174.12/issuetoc.

Treatment of Alzheimer disease using combination therapy with plasma exchange and haemapheresis with albumin and intravenous immunoglobulin: Rationale and treatment approach of the AMBAR (Alzheimer Management By Albumin Replacement) study

IntroductionThere is a growing interest in new therapeutic strategies for the treatment of Alzheimer disease (AD) which focus on reducing the beta-amyloid peptide (Aβ) burden in the brain by sequestering plasma Aβ, a large proportion of which is bound to albumin and other proteins. This review discusses the concepts of interaction between Aβ and albumin that have given rise to AMBAR (Alzheimer's Disease Management by Albumin Replacement) project, a new multicentre, randomised, controlled clinical trial for the treatment of AD. DevelopmentResults from preliminary research suggest that Albutein® (therapeutic albumin, Grifols) contains no quantifiable levels of Aβ. Studies also show that Albutein® has Aβ binding capacity. On the other hand, AD entails a high level of nitro-oxidative stress associated with fibrillar aggregates of Aβ that can induce albumin modification, thus affecting its biological functions. Results from the phase ii study confirm that using therapeutic apheresis to replace endogenous albumin with Albutein® 5% is feasible and safe in patients with AD. This process resulted in mobilisation of Aβ and cognitive improvement in treated patients. The AMBAR study will test combination therapy with therapeutic apheresis and haemopheresis with the possible leverage effect of Albutein® with intravenous immunoglobulin replacement (Flebogamma® DIF). Cognitive, functional, and behavioural changes in patients with mild to moderate AD will be assessed. ConclusionsThe AMBAR study represents a new therapeutic perspective for AD.

Nitroglycerin induces DNA damage and vascular cell death in the setting of nitrate tolerance

Nitroglycerin (GTN) and other organic nitrates are widely used vasodilators. Their side effects are development of nitrate tolerance and endothelial dysfunction. Given the potential of GTN to induce nitro-oxidative stress, we investigated the interaction between nitro-oxidative DNA damage and vascular dysfunction in experimental nitrate tolerance. Cultured endothelial hybridoma cells (EA.hy 926) and Wistar rats were treated with GTN (ex vivo: 10-1000µM; in vivo: 10, 20 and 50mg/kg/day for 3days, s.c.). The level of DNA strand breaks, 8-oxoguanine and O (6)-methylguanine DNA adducts was determined by Comet assay, dot blot and immunohistochemistry. Vascular function was determined by isometric tension recording. DNA adducts and strand breaks were induced by GTN in cells in vitro in a concentration-dependent manner. GTN in vivo administration leads to endothelial dysfunction, nitrate tolerance, aortic and cardiac oxidative stress, formation of DNA adducts, stabilization of p53 and apoptotic death of vascular cells in a dose-dependent fashion. Mice lacking O (6)-methylguanine-DNA methyltransferase displayed more vascular O (6)-methylguanine adducts and oxidative stress under GTN therapy than wild-type mice. Although we were not able to prove a causal role of DNA damage in the etiology of nitrate tolerance, the finding of GTN-induced DNA damage such as the mutagenic and toxic adduct O (6)-methylguanine, and cell death supports the notion that GTN based therapy may provoke adverse side effects, including endothelial function. Further studies are warranted to clarify whether GTN pro-apoptotic effects are related to an impaired recovery of patients upon myocardial infarction.

Canavanine Alters ROS/RNS Level and Leads to Post-translational Modification of Proteins in Roots of Tomato Seedlings

Canavanine (CAN), a structural analog of arginine (Arg), is used as a selective inhibitor of inducible NOS in mammals. CAN is incorporated into proteins’ structure in the place of Arg, leading to the formation of aberrant compounds. This non-protein amino acid is found in legumes, e.g., Canavalia ensiformis (L.) DC. or Sutherlandia frutescens (L.) R.Br. and acts as a strong toxin against herbivores or plants. Tomato (Solanum lycopersicum L.) seedlings were treated for 24–72 h with CAN (10 or 50 μM) inhibiting root growth by 50 or 100%, without lethal effect. We determined ROS level/production in root extracts, fluorescence of DAF-FM and APF derivatives corresponding to RNS level in roots of tomato seedlings and linked CAN-induced restriction of root growth to the post-translational modifications (PTMs) of proteins: carbonylation and nitration. Both PTMs are stable markers of nitro-oxidative stress, regarded as the plant’s secondary response to phytotoxins. CAN enhanced H2O2 content and superoxide radicals generation in extracts of tomato roots and stimulated formation of protein carbonyl groups. An elevated level of carbonylated proteins was characteristic for the plants after 72 h of the culture, mainly for the roots exposed to 10 μM CAN. The proteolytic activity was stimulated by tested non-protein amino acid. CAN treatment led to decline of fluorescence of DAF-FM derivatives, and transiently stimulated fluorescence of APF derivatives. Short-term exposure of tomato seedlings to CAN lowered the protein nitration level. Activity of peroxidase, polyamine oxidase and NADPH oxidase, enzymes acting as modulators of H2O2 concentration and governing root architecture and growth were determined. Activities of all enzymes were stimulated by CAN, but no strict CAN concentration dependence was observed. We conclude, that although CAN treatment led to a decline in the nitric oxide level, PTMs observed in roots of plants exposed to CAN are linked rather to the formation of carbonyl groups than to nitration, and are detected particularly after 24 h. Thus, oxidative stress and oxidative modifications of proteins seems to be of significant importance in the rapid response of plants to CAN.

Association between low thyroid-stimulating hormone, posterior cortical atrophy and nitro-oxidative stress in elderly patients with cognitive dysfunction.

IntroductionCortical atrophy is known to be a valuable sign of cognitive decline. The purpose of this study was to assess the association between low thyroid-stimulating hormone (TSH), posterior cortical atrophy (Koedam score – KS) and nitro-oxidative stress in elderly patients.Material and methodsA study (SG) and a control group (CG), each subdivided by gender, were investigated. Subjects older than 59 years with low serum TSH level and with mild cognitive impairment were included in the SG. The CG was formed by subjects free of significant cortical atrophy and free or thyroid dysfunction. Demographic and clinical characteristics of the patients (Mini Mental State Examination, MMSE), Koedam score on cranial magnetic resonance imaging, and blood parameters (TSH, FT4, and nitric oxide – NOx) were assessed.ResultsSubjects in the study group had fewer years of education above the 8th grade compared with the control group (p < 0.0001). A significantly higher percentage of subjects in the study group had a Koedam score of 2 or 3 compared with controls, who had in the majority of cases a Koedam score of zero (p < 0.02). Significantly higher NOx levels were observed when study groups of both genders were compared with corresponding controls (p < 0.001). No significant differences were observed with regard to FT4 (p > 0.70). Nitric oxide was found to be significantly associated with TSH (p < 0.03) and KS (p < 0.002) when the whole study group was considered as well as when just the non-smoker study group was investigated.ConclusionsOur study revealed an association between subclinical thyroid hypofunction, nitro-oxidative stress, and posterior cortical atrophy as an early stage of global atrophy.

New Insight into 2-Methoxyestradiol- a Possible Physiological Link between Neurodegeneration and Cancer Cell Death.

The pathways leading to development of neurodegenerative disease and cancer cell death may overlap. Interestingly, the main neurodegeneration-causing factors have been suggested to play a pivotal role in the regulation of tumor growth. Another common component in neurodegenerations and cancer is nitro-oxidative stress. Depending on the environmental conditions, the nitro-oxidative stress may lead to cancerogenesis or cancer cell death. On the other side, it may also result in a variety of neurodegenerative diseases. Based on the literature data, we suggest that 2-methoxyestradiol may be considered as a physiological link between development of neurodegenerations and cancer cell death. 2-methoxyestradiol is a physiological compound, a metabolite of 17β-estradiol. Interestingly, 2-methoxyestradiol, branded as Panzem, is also considered as a potent anticancer agent, currently investigated in several clinical trials. The review aims on making closer the facts concerning anticancer and plausible neurotoxic activities of 2-methoxyestradiol, which may suggest adverse neurological long-term effects of the drug in cancer patients.

Exercise does not activate the β3 adrenergic receptor–eNOS pathway, but reduces inducible NOS expression to protect the heart of obese diabetic mice

Obesity and diabetes are associated with higher cardiac vulnerability to ischemia-reperfusion (IR). The cardioprotective effect of regular exercise has been attributed to β3-adrenergic receptor (β3AR) stimulation and increased endothelial nitric oxide synthase (eNOS) activation. Here, we evaluated the role of the β3AR-eNOS pathway and NOS isoforms in exercise-induced cardioprotection of C57Bl6 mice fed with high fat and sucrose diet (HFS) for 12weeks and subjected or not to exercise training during the last 4 weeks (HFS-Ex). HFS animals were more sensitive to in vivo and ex vivo IR injuries than control (normal diet) and HFS-Ex mice. Cardioprotection in HFS-Ex mice was not associated with increased myocardial eNOS activation and NO metabolites storage, possibly due to the β3AR-eNOS pathway functional loss in their heart. Indeed, a selective β3AR agonist (BRL37344) increased eNOS activation and had a protective effect against IR in control, but not in HFS hearts. Moreover, iNOS expression, nitro-oxidative stress (protein s-nitrosylation and nitrotyrosination) and ROS production during early reperfusion were increased in HFS, but not in control mice. Exercise normalized iNOS level and reduced protein s-nitrosylation, nitrotyrosination and ROS production in HFS-Ex hearts during early reperfusion. The iNOS inhibitor 1400W reduced in vivo infarct size in HFS mice to control levels, supporting the potential role of iNOS normalization in the cardioprotective effects of exercise training in HFS-Ex mice. Although the β3AR-eNOS pathway is defective in the heart of HFS mice, regular exercise can protect their heart against IR by reducing iNOS expression and nitro-oxidative stress.

Oral treatment with a zinc complex of acetylsalicylic acid prevents diabetic cardiomyopathy in a rat model of type-2 diabetes: activation of the Akt pathway.

BackgroundType-2 diabetics have an increased risk of cardiomyopathy, and heart failure is a major cause of death among these patients. Growing evidence indicates that proinflammatory cytokines may induce the development of insulin resistance, and that anti-inflammatory medications may reverse this process. We investigated the effects of the oral administration of zinc and acetylsalicylic acid, in the form of bis(aspirinato)zinc(II)-complex Zn(ASA)2, on different aspects of cardiac damage in Zucker diabetic fatty (ZDF) rats, an experimental model of type-2 diabetic cardiomyopathy.MethodsNondiabetic control (ZL) and ZDF rats were treated orally with vehicle or Zn(ASA)2 for 24 days. At the age of 29–30 weeks, the electrical activities, left-ventricular functional parameters and left-ventricular wall thicknesses were assessed. Nitrotyrosine immunohistochemistry, TUNEL-assay, and hematoxylin-eosin staining were performed. The protein expression of the insulin-receptor and PI3K/AKT pathway were quantified by Western blot.ResultsZn(ASA)2-treatment significantly decreased plasma glucose concentration in ZDF rats (39.0 ± 3.6 vs 49.4 ± 2.8 mM, P < 0.05) while serum insulin-levels were similar among the groups. Data from cardiac catheterization showed that Zn(ASA)2 normalized the increased left-ventricular diastolic stiffness (end-diastolic pressure–volume relationship: 0.064 ± 0.008 vs 0.084 ± 0.014 mmHg/µl; end-diastolic pressure: 6.5 ± 0.6 vs 7.9 ± 0.7 mmHg, P < 0.05). Furthermore, ECG-recordings revealed a restoration of prolonged QT-intervals (63 ± 3 vs 83 ± 4 ms, P < 0.05) with Zn(ASA)2. Left-ventricular wall thickness, assessed by echocardiography, did not differ among the groups. However histological examination revealed an increase in the cardiomyocytes’ transverse cross-section area in ZDF compared to the ZL rats, which was significantly decreased after Zn(ASA)2-treatment. Additionally, a significant fibrotic remodeling was observed in the diabetic rats compared to ZL rats, and Zn(ASA)2-administered ZDF rats showed a similar collagen content as ZL animals. In diabetic hearts Zn(ASA)2 significantly decreased DNA-fragmentation, and nitro-oxidative stress, and up-regulated myocardial phosphorylated-AKT/AKT protein expression. Zn(ASA)2 reduced cardiomyocyte death in a cellular model of oxidative stress. Zn(ASA)2 had no effects on altered myocardial CD36, GLUT-4, and PI3K protein expression.ConclusionsWe demonstrated that treatment of type-2 diabetic rats with Zn(ASA)2 reduced plasma glucose-levels and prevented diabetic cardiomyopathy. The increased myocardial AKT activation could, in part, help to explain the cardioprotective effects of Zn(ASA)2. The oral administration of Zn(ASA)2 may have therapeutic potential, aiming to prevent/treat cardiac complications in type-2 diabetic patients.

Medical ozone therapy reduces shock wave therapy-induced renal injury

Objectives: Extracorporeal shock wave (ESW) lithotripsy is the preferred treatment modality for uncomplicated kidney stones. More recently free oxygen radical production following ESW application has been considered to be crucial in shock wave-induced renal damage. It has been shown that ozone therapy (OT) has ameliorative and preventive effects against various pathological conditions due to increased nitro-oxidative stress. In current study, we aimed to evaluate the efficacy of OT against ESW-induced renal injury. Methods: Twenty-four male Sprague–Dawley rats were divided into three groups: sham-operated, ESW, and ESW + OT groups. All groups except sham-operated group were subjected to ESW procedure. ESW + OT group received 1 mg/kg/day of oxygen/ozone mixture intraperitoneally at 2 h before ESW, and OT was continued once a day for consecutive three days. The animals were killed at the 4th day, and kidney tissue and blood samples were harvested for biochemical and histopathologic analysis. Results: Serum ALT and AST levels, serum neopterin, tissue nitrite/nitrate levels, and tissue oxidative stress parameters were increased in the ESW group and almost came close to control values in the treatment group (p < 0.05, ESW vs. ESW + OT). Histopathological injury scores were significantly lower in treatment group than the ESW group (p < 0.05, ESW vs. ESW + OT). Immunohistochemical iNOS staining scores in ESW group were higher than those of sham-operated group (p < 0.05, ESW vs. sham-operated), iNOS staining scores in OT group were significantly lower than the ESW group (p < 0.05, ESW + OT vs. ESW). Conclusion: OT ameliorates nitro-oxidative stress and reduces the severity of pathological changes in the experimental ESW-induced renal injury of rat model.

Effect of maximal-intensity exercise on systemic nitro-oxidative stress in men and women

Objectives: The aim of this study was to test the hypotheses: (1) there is a negative correlation between protein and lipid oxidative damage following maximal-intensity exercise, and oxygen uptake and work intensity (%VO2max) at the respiratory compensation point (RCP) in women and men; (2) nitro-oxidative stress following maximal-intensity exercise results from the intensification of anaerobic processes and muscle fibre micro-damage.Methods: Study participants comprised 20 women (21.34±1.57 years) and 20 men (21.97±1.41 years) who performed a treadmill incremental test (IT); VO2max: 45.08 ± 0.91 and 57.38 ± 1.22 mL kg−1 min−1 for women and men, respectively. The oxidized low-density lipoprotein (ox-LDL), 3-nitrotyrosine (3-NT) concentration and creatine kinase (CK) as well as lactate dehydrogenase (LDH) activity were measured in the blood serum, and total antioxidative capacity (TAC) and lactate concentration (Lac) were determined in blood plasma before and after IT.Results: After the IT, increases in ox-LDL, 3-NT, CK, and LDH were seen in both groups (P < 0.05). After the IT, an increase in the TAC was only observed in women (P < 0.05). The post-exercise-induced increase in Lac was significantly higher in men than in women. Only in the group of women was a positive correlation (P < 0.05) between the post-exercise increase in TAC and changes in CK activity and LDH found.Conclusions: The gain of ox-LDL and 3-NT following maximal-intensity exercise is independent of VO2max, oxygen consumption and exercise intensity at RCP. This increase of ox-LDL and 3-NT is indicative of similar lipid and protein damage in women and men. A significant increase in TAC in women following maximal-intensity exercise is the result of muscle fibre micro-injuries.

Chrysin, a PPAR-γ agonist improves myocardial injury in diabetic rats through inhibiting AGE-RAGE mediated oxidative stress and inflammation

AGE-RAGE interaction mediated oxidative stress and inflammation is the key mechanism involved in the pathogenesis of cardiovascular disease in diabetes. Inhibition of AGE-RAGE axis by several PPAR-γ agonists has shown positive results in ameliorating cardio-metabolic disease conditions. Chrysin, a natural flavonoid has shown to possess PPAR-γ agonist activity along with antioxidant and anti-inflammatory effect. Therefore, the present study was designed to evaluate the effect of chrysin in isoproterenol-induced myocardial injury in diabetic rats. In male albino Wistar rats, diabetes was induced by single injection of streptozotocin (70 mg/kg, i.p.). After confirmation of the diabetes, rats were treated with vehicle (1.5 mL/kg, p.o.), chrysin (60 mg/kg, p.o.) or PPAR-γ antagonist GW9662 (1 mg/kg, i.p.) for 28 days. Simultaneously, on 27th and 28th day myocardial injury was induced by isoproterenol (85 mg/kg, s.c.). Chrysin significantly ameliorated cardiac dysfunction as reflected by improved MAP, ±LVdP/dtmax and LVEDP in diabetic rats. This improvement was associated with increased PPAR-γ expression and reduced RAGE expression in diabetic rats. Chrysin significantly decreased inflammation through inhibiting NF-κBp65/IKK-β expression and TNF-α level. Additionally, chrysin significantly reduced apoptosis as indicated by augmented Bcl-2 expression and decreased Bax and caspase-3 expressions. Furthermore, chrysin inhibited nitro-oxidative stress by normalizing the alteration in 8-OHdG, GSH, TBARS, NO and CAT levels and Nox4, MnSOD, eNOS and NT expressions. Co-administration of GW9662 significantly blunted the chrysin mediated cardioprotective effect as there was increase in oxidative stress, inflammation and apoptosis markers. Chrysin significantly ameliorated isoproterenol-induced myocardial injury in diabetic rats via PPAR-γ activation and inhibition of AGE-RAGE mediated oxidative stress and inflammation.