ABSTRACT
Objetives: The goal of this study was to determine if systemic and peritoneal oxidative stress biomarkers are related to each other and to retrograde menstruation in endometriosis. Methods: Plasma and peritoneal fluid oxidative stress biomarkers and hemoglobin and erythrocytes in peritoneal fluid as retrograde menstruation indicators, were measured in 28 patients with endometriosis and 23 without endometriosis. Results: In the peritoneal fluid, carbonyls and lipohydroperoxides, indicative of protein and lipid oxidative damage, were higher in endometriosis group (21%, p = 0.016 and 46%, p = 0.009, respectively). However, these biomarkers were not different in the blood plasma of both groups, and only protein dityrosine, was increased in the plasma of endometriosis group (31%, p = 0.04). The peritoneal fluid hemoglobin content was not higher in the endometriosis group, nor related to carbonyls and lipohydroperoxides. Additionally, the peritoneal fluid oxidative biomarkers were not correlated with the blood plasma ones, and only malondialdehyde, and ischemia-modified albumin were almost two times higher in peritoneal fluid. Discussion: Our results show a peritoneal and systemic oxidative stress biomarkers increase in endometriosis, but not related to each other, and do not support the hypothesis of an increase in hemoglobin-iron supply towards the peritoneal cavity that causes oxidative damage.
Subject(s)
Biomarkers/metabolism , Endometriosis/metabolism , Oxidative Stress/physiology , Adolescent , Adult , Ascitic Fluid/metabolism , Female , Humans , Serum Albumin, Human/metabolism , Young AdultABSTRACT
During Chronic Obstructive Pulmonary Disease (COPD) progression, the intracellular antioxidant defence in RBCs must preserve the integrity of the plasmalemma through NADPH+ generation to obtain a sufficient number of reduced non-protein SH-groups. Here, we studied the activities of enzymes in RBCs that are related to glutathione metabolism under conditions of increasing oxidative stress, which are associated with COPD progression, by increasing cellular damage in vitro with PM2.5, a ROS generator. The study included 43 patients, who were separated according to their GOLD classification into moderate and severe groups, along with 11 healthy volunteers (HV). Blood samples were analysed for G6PD, GAPDH, GPx, and GR. The results showed significant decreases in the oxidation of the G6PD, GR and GPx proteins, resulting in decreased enzymatic activity. By contrast, an increase (p<0.05) in GAPDH was observed, suggesting a pool of ATP on the membrane. However, it is evident that RBCs are damaged during the progression of COPD, although their integrity is preserved, and they retain limited function, thus allowing patient survival without haemolysis.
Subject(s)
Erythrocytes/drug effects , Erythrocytes/enzymology , Glyceraldehyde-3-Phosphate Dehydrogenases/metabolism , Particulate Matter/toxicity , Pulmonary Disease, Chronic Obstructive/blood , Pulmonary Disease, Chronic Obstructive/enzymology , Case-Control Studies , Cohort Studies , Disease Progression , Enzyme Activation , Erythrocyte Membrane/drug effects , Erythrocyte Membrane/enzymology , Erythrocyte Membrane/metabolism , Erythrocytes/metabolism , Glutathione Peroxidase/blood , Glutathione Reductase/blood , Glyceraldehyde-3-Phosphate Dehydrogenases/blood , Hemolysis , Humans , Oxidation-Reduction , Oxidative Stress/drug effects , Particle Size , Particulate Matter/blood , Pulmonary Disease, Chronic Obstructive/metabolism , Reactive Oxygen Species/metabolism , Urban PopulationABSTRACT
Particulate matters (PM) produce adverse effects on the respiratory system and cause COPD. These effects are thought to involve intrinsic generation of ROS which are present in ambient PM (transition metals and aromatic organic compounds). Here, we examined the chemical composition and ultra-microscopic structure of PM2.5. The effect of this PM was studied in red blood cell (RBC) membranes (ghosts) from healthy volunteers (n = 11) and COPD patients (n = 43). These effects were compared with that produced by a Fenton metal-catalytic ROS generator. Oxidative biomarkers and cell damage were singificantly increased in presence of PM2.5 or ROS generator in RBC of COPD patients as compared with those in cells from healthy volunteers. In contrast, total SH groups, band 3 phospho-tyrosine phosphatase (PTPase) and glucose-6 phosphate dehydrogenase (G6PD) activities were all diminished in cells from COPD patients. In conclusion, PM2.5 increases damage to RBCs from COPD patients, decreases the activity of PTPase and G6PD, and alters the function of the anionic exchanger (AE1) and the antioxidant response by decreasing SH groups.
Subject(s)
Air Pollutants/toxicity , Erythrocytes/drug effects , Pulmonary Disease, Chronic Obstructive/metabolism , Reactive Oxygen Species/metabolism , Urban Health , Aged , Female , Humans , Male , Middle Aged , Particle Size , Pulmonary Disease, Chronic Obstructive/bloodABSTRACT
The change produced by oxidative stress on proteins (cross-links, backbone cleavage, amino acid modification) generates structural changes with a wide range of consequences such as increased propensity to the aggregation or proteolysis, altered immunogenicity and frequently enzymatic and binding inhibition. Insulin is particularly sensitive to conformational changes, aggregation and cross-linking; any change on insulin could impair its function. We have examined the biological activity of insulin modified by hydroxyl radical and exposed to acrolein in rats and adiposites. We found out important changes that we have shown as prototype of possible effect of oxidative stress on the structural and functional damage to insulin. Whereas, hydroxyl radical and acrolein both have diminished the hypoglycemic effect of insulin in vivo, and the effect of acrolein seems be to involved in carbonylation and not derived from inter-molecular cross-links formation or aggregates. The effect was highly stimulated at alkaline pH, concomitant with carbonyl formation and then probably aldolic condensation type reaction-dependent. Hydroxyls radical generates tyrosine derivative formation and introduces non aldehyde dependent carbonyls in the insulin molecule.
Subject(s)
Insulin/chemistry , Insulin/metabolism , Oxidative Stress/drug effects , 3T3-L1 Cells , Acrolein/pharmacology , Animals , Carbohydrate Metabolism/drug effects , Glucose/metabolism , Hydroxyl Radical/pharmacology , Mice , Protein Carbonylation/drug effects , RatsABSTRACT
Oxidative stress damage to biomolecules has been implicated in several diseases including diabetes mellitus. In the present study, we investigated the effect of oxidative stress in whole blood (WB) from diabetic patients (n = 60) on recombinant human insulin. Insulin was incubated with WB obtained from diabetic patients (DP) who had hyperglycemia (>300 mg/dL) or from 41 healthy volunteers (HV). Whole blood of DP, unlike WB of HV, induced higher values of formazan (142%), dityrosines (279%), and carbonyls (58%) in the insulin residues. Interestingly, the insulin modified by WB of DP showed less hypoglycemic activity in rat (30%) in comparison with insulin incubated with WB of HV. The incubation of insulin in WB from DP induces chemical changes in insulin and a decrease in its biological activity, events that might be associated with the high levels of oxidative stress markers found in the plasma of these patients.
Subject(s)
Diabetes Mellitus, Type 2/blood , Insulin/metabolism , Oxidative Stress/physiology , Area Under Curve , Biomarkers , Blood Glucose/metabolism , Female , Formazans/metabolism , Humans , Hypoglycemic Agents/therapeutic use , Injections, Intraperitoneal , Insulin/chemistry , Iron/blood , Male , Malondialdehyde/metabolism , Middle Aged , Oxidation-Reduction , Protein Carbonylation/drug effects , Reactive Oxygen Species/metabolism , Tyrosine/analogs & derivatives , Tyrosine/metabolismABSTRACT
In myocardial infarctions with ST-segment elevation, ischemia followed by reperfusion (IR) leads to arrhythmia, myocardial stunning and endothelial dysfunction injury by reactive oxygen species (ROS). To determine the impact of ROS, we examined the effect of antioxidant vitamins on biochemical changes and arrhythmias induced by reperfusion before and after therapeutic thrombolysis (Actilyse). As compared with those receiving placebo, in individuals who received antioxidants, there was a significant decrease in premature ventricular beats (100% vs 38%), atrial fibrillation (44% vs 6%), ventricular tachycardia (31% vs 0%), first-degree atrial-ventricular block (44% vs 6%), plasma malondialdehyde at the first hour after initiation of thrombolysis (1.07 +/- 0.10 vs 0.53 +/- 0.10 nmols plasma malondialdehyde/mg protein) and circulating neutrophils after 24 hr after reperfusion. The antioxidant capacity of plasma was increased from 1.89 +/- 0.15 to 3.00 +/- 0.31 units/mg protein and paraoxonase-1 rose from 0.77 +/- 0.08 to 1.27 +/- 0.11 nmol/min/mg protein. These findings suggest that antioxidants might be useful as adjuvants in controlling reperfusion induced arrhythmias following therapeutic alteplase thrombolysis.
Subject(s)
Antioxidants/therapeutic use , Arrhythmias, Cardiac/drug therapy , Myocardial Infarction/drug therapy , Myocardial Reperfusion Injury/complications , Arrhythmias, Cardiac/etiology , Biomarkers/metabolism , Coronary Angiography , Electrocardiography , Female , Humans , Male , Middle Aged , Minerals/therapeutic use , Myocardial Infarction/complications , Myocardial Infarction/diagnosis , Ventricular Function, Left , Vitamins/therapeutic useABSTRACT
The formation of dityrosine of human insulin oxidized by metal-catalyzed oxidation system (H2O2/Cu) was estimated by fluorescent methods. The oxidation of tyrosine and phenylalanine residues present on the insulin molecule was evident after 2 minutes of in vitro oxidation due to the formation of protein-bound dityrosine. The success of oxidative protein modification was followed until available aromatic residues were consumed (60 minutes), measured by their emission at 405 nm. The structural and chemical changes on insulin molecule are related to the loss of biological activity as assessed by measuring the increase of U-14C-glucose utilization by human adipose tissue in a radiorespirometry system. The oxidation of glucose (14CO2 production) of the adipose cells was increased 35 % (301 +/- 119 to 407 +/- 182 cpm/mg in dry weight. P < 0.05) in presence of 0.1 IU and 69 % (301 +/- 119 to 510 +/- 266 cpm/dry weight. P < 0.05) for 1.0 IU of insulin. The recombinant human insulin oxidized for 5 minutes only increased the glucose oxidation by 25 %. In conclusion, these observations show that dityrosine formation and other oxidative chemical changes of insulin due to its in vitro oxidation decrease and can abolish its biological activity.
Subject(s)
Adipose Tissue/metabolism , Glucose/metabolism , Insulin/metabolism , Reactive Oxygen Species/metabolism , Tyrosine/analogs & derivatives , Carbon Radioisotopes , Humans , Insulin/chemistry , Oxidation-Reduction , Protein Conformation , Tyrosine/analysisABSTRACT
Oxidative stress occurs when the production of reactive oxygen species (ROS) exceeds the endogenous antioxidant defense. Peroxidations induced by ROS are the key of chemical and structural modifications of biomolecules including circulating proteins. To elucidate the effect of ROS on circulating proteins and considering the presence of oxidative stress in Diabetes Mellitus, the effects of ROS, in vitro, on human insulin were studied. We utilized the Fenton reaction for free hydroxyl radical (HO*) generation in presence of human recombinant insulin measuring chemical changes on its molecular structure. The induced changes in insulin were: a) significant increase on absorbance (280 nm) due to phenylalanine hydroxylation (0.023 +/- 0.007 to 0.13 +/- 0.07). b) Peroxidation products formed on amino acids side branches (peroxyl and alcohoxyl group); measured as increased capacity of reduce nitroblue of tetrazolium (NBT) to formazan (0.007 +/- 0.007 to 0.06 +/- 0.02). c) Increased concentration of free carbonyl groups (8.8 +/- 8.7 to 45.6 +/- 20.2 pmoles dinitrophenylhidrazones/nmol insulin) with lost of secondary structure, and d) Modification of epithopes decreasing the insulin antigen-antibody reactivity measured as a decrease in insulin concentration by RIA. In conclusion, the radical hydroxyl in vitro is able to induce molecular modifications on insulin.
Subject(s)
Insulin/chemistry , Insulin/metabolism , Reactive Oxygen Species , Carbon/chemistry , Diabetes Mellitus/metabolism , Epitopes/chemistry , Formazans/chemistry , Humans , Hydroxyl Radical , Nitroblue Tetrazolium/chemistry , Oxidative Stress , Phenylalanine/chemistry , RadioimmunoassayABSTRACT
En el endometrio el citoesqueleto participa en todas las funciones mecánicas de la célula, en el movimiento y reacomodo de organelos y proteínas solubles y en el metabolismo en general. El epitelio endometrial por su morfología y aparente homogeneidad celular, se ha estudiado más que el estroma. Se sabe que los filamentos intermedios muestran un patrón característico típico de la clase celular. Durante la preñez y la pseudopreñez, en la región apical de las células epiteliales tanto luminales como glandulares predomina la queratina sobre la región basolateral, en tanto que la vimetina solo se encuentra en las células epiteliales luminales, y se incrementa el día de la implantación. En humanos y roedores, la desmina solo se expresa en la decidua. Se piensa que los filamentos intermedios, participan en la difusión de proteínas de membrana cambiando la polaridad. Los microfilamentos intervienen en la regulación de la forma y movilidad celular. En el epitelio luminal participan en las transformaciones de la superficie uterina, como son las microvellosidades. Al sistema de microtúbulos (MT) en el endometrio y otros órganos se le ha relacionado con la psisción y movimiento de orgánulos como son los lisosomas, mitocondrias o aparato de Golgio; además se ha demostrado que los MT también intervienen en la síntesis de DNA, ya que drogas como la colchicina impiden estos fenómenos