RESUMEN
Our study concerns the effects of exposure to lead chloride on the morphology, K(+) efflux, SO(4)(-) influx and GSH levels of the human erythrocyte. Blood was collected in heparinized tubes and washed three times. The cells were suspended at 3% hematocrit and incubated for 1 h at 25°C in a medium containing increasing concentrations of lead chloride (0, 0.3, 0.5 and 1 µM). After incubation, the suspensions were centrifuged and the erythrocyte pellets were divided into three aliquots for testing. The results show: an increase in the permeability of erythrocytes treated with lead chloride with consequent damage and cellular death, especially in the presence of high concentrations; an increase in potassium ion efflux; alterations in the morphology and membrane structure of the red blood cells; and a decrease in sulphate uptake, due either to the oxidative effect of this compound on the band 3 protein, which loses its biological valence as a carrier of sulphate ions, or to a decrease in the ATP erythrocyte concentration. In conclusion, the exposure of erythrocytes to Pb(2+) ions leads to a reduction in the average lifetime of the erythrocytes and the subsequent development of anemia. These data are discussed in terms of the possible effect of lead on the reduction-oxidation systems of the cell. Oxidant agents, such as lead, are known to cross-link integral membrane proteins, leading to K/Cl-cotransport. The increased K(+) efflux affects the altered redox state.
Asunto(s)
Membrana Eritrocítica/efectos de los fármacos , Glutatión/metabolismo , Plomo/toxicidad , Sulfatos/metabolismo , Adenosina Trifosfato/metabolismo , Proteína 1 de Intercambio de Anión de Eritrocito/metabolismo , Aniones/química , Apoptosis/efectos de los fármacos , Permeabilidad de la Membrana Celular/efectos de los fármacos , Membrana Eritrocítica/metabolismo , Humanos , Cinética , Oxidación-Reducción , Potasio/metabolismoRESUMEN
The objective of this study was to assess the effects of nickel chloride on human and rainbow trout erythrocytes in vitro. The cells were incubated with 0, 0.5 and 1 mM nickel chloride for 1 h at pH 7.40 and 25 degrees C, then K(+) efflux, SO (4) (2-) uptake and GSH and GSSG concentrations were measured. In both kind of cells, "high concentration" nickel treatment increased KCl efflux with respect to the control. The SO (4) (2-) uptake was not significantly different at "low nickel concentration" but was lower in erythrocytes treated with 1 mM nickel chloride; the rate constant of SO (4) (2-) uptake decreased by 35% in human erythrocytes and by 44% in fish erythrocytes. Nickel chloride also acts on cellular metabolism and in particular on erythrocyte glutathione peroxidase with consequent increase in oxidative stress; the data show a significant decrease in intracellular GSH in both human (25%) and fish erythrocytes (18%) after treatment with nickel chloride, with concomitantly high GSSG concentrations and lower GSH/GSSG ratios.
Asunto(s)
Proteína 1 de Intercambio de Anión de Eritrocito/metabolismo , Eritrocitos/efectos de los fármacos , Níquel/farmacología , Animales , Tamaño de la Célula , Glutatión/sangre , Humanos , Oncorhynchus mykiss , Oxidación-Reducción , Estrés Oxidativo , Potasio/metabolismo , Sulfatos/metabolismoRESUMEN
L-Arginine is an essential amino acid for infants and growing children, as well as for pregnant women. This amino acid is a substrate for at least 5 enzymes identified in mammals, including arginase, arginine-glycine transaminase, kyotorphine synthase, nitric oxide synthase, and arginine decarboxylase. L-arginine is essential for the synthesis of creatine, urea, polyamines, nitric oxide, and agmatine. Arginine may be considered an essential amino acid in sepsis, and its supplementation could be beneficial in this clinical setting by improving microcirculation and protein anabolism. Rats receiving arginine-supplemented parenteral nutrition showed an increased ability to synthesize acute phase proteins when challenged with sepsis. Finally, L-arginine exerts antihypertensive and antiproliferative effects on vascular smooth muscles. It has been shown to reduce systemic blood pressure in some forms of experimental hypertension. Endothelial dysfunction and reduced nitric oxide bioactivity are associated with increased incidence of cardiovascular diseases. A beneficial effect of acute and chronic L-arginine supplementation on endothelial derived nitric oxide production and endothelial function has been shown. In end-stage renal disease patients, the rate of de novo arginine synthesis seemed to be preserved. Our preliminary data on a group of dialysis patients showed that predialysis arginine levels were stable in a normal range during the dialysis session and that hypertensive patients had lower arginine-citrulline ratio than normotensive patients.
Asunto(s)
Arginina/uso terapéutico , Fallo Renal Crónico/terapia , Diálisis Renal , Adulto , Anciano , Animales , Arginina/análogos & derivados , Arginina/sangre , Arginina/fisiología , Citrulina/sangre , Nefropatías Diabéticas/tratamiento farmacológico , Endotelio Vascular/fisiopatología , Femenino , Humanos , Hipertensión , Fallo Renal Crónico/fisiopatología , Masculino , Persona de Mediana Edad , Óxido Nítrico/fisiologíaRESUMEN
Inducible nitric oxide synthase knock-out (iNOS(-/-)) mice are valid models of investigation for the role of iNOS in patho-physiological conditions. There are no available data concerning neuroactive amino acid levels of iNOS(-/-) mice and their behaviour in response to pentylenetetrazole (PTZ). We found no significant differences in the convulsive dose 50 (CD(50)) between iNOS(-/-) and control (iNOS(+/+)) mice, however, iNOS(-/-) mice reach the kindled status more slowly than control, suggesting that in basal condition the GABA-benzodiazepine inhibitory inputs are unaltered by iNOS mutation. Clear differences between iNOS(+/+) and iNOS(-/-) mice amino acid concentrations were evident both in basal conditions and after kindling. Our results show that aspartate was significantly lower in all brain areas studied except the brain stem whereas glutamate and glutamine were significantly higher in the cortex, hippocampus and brain stem. GABA was slightly and not significantly higher in the cortex, hippocampus and brain stem, whereas taurine was significantly higher in all areas except diencephalon and glycine was significantly lower in the diencephalon and cerebellum. In this context, the inability of iNOS(-/-) mice to increase the NO levels following PTZ administrations indicate that NO might play a pro-epileptogenic role in the genesis and development of some types of epilepsy. Since there is no correlation between neurotransmitter levels and the development of kindling, it is possible to exclude that the difference between the two strains is due to an imbalance between the considered neurotransmitters, and it is then possible that this difference is due to the presence of iNOS, which might be involved in long term plasticity of the brain.
Asunto(s)
Aminoácidos/metabolismo , Encéfalo/enzimología , Antagonistas del GABA/farmacología , Excitación Neurológica , Óxido Nítrico Sintasa de Tipo II/metabolismo , Animales , Encéfalo/fisiopatología , Convulsivantes , Antagonistas de Receptores de GABA-A , Ratones , Ratones Noqueados , Óxido Nítrico Sintasa de Tipo II/genética , Pentilenotetrazol , Convulsiones/tratamiento farmacológico , Convulsiones/fisiopatología , Taurina/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Genetic animal models have contributed significantly to our understanding of epilepsy causes. Lethargic mice are considered a valid model of absence epilepsy, which have been shown to possess behavioral, electrographic and pharmacological profiles similar to those of humans with absence epilepsies. Single gene mutations that comprise the beta4 subunit of voltage-sensitive Ca2+ channels underlie the spontaneous discharges of the absence, non-convulsive seizures of lethargic mice. There are no available data concerning how the mutant channels actually behave at terminals in response to chemical activation by subconvulsant stimulation with pentylenetetrazole. In this study, we found no significant difference in the convulsive dose 50 between lethargic and control mice. Lethargic mice showed a more rapid development of kindling to pentylenetetrazole than control animals. No significant differences were observed between the groups of mice rechallenged with pentylenetetrazole 30 or 60 days after the end of the chronic treatment. Marked differences in brain amino acid levels were found between the two strains of mice in basal conditions and after kindling. In conclusion, our results indicate that lethargic mice show a range of biochemical and behavioral changes, correlated in particular with a higher susceptibility to develop kindled seizures.
Asunto(s)
Aminoácidos/metabolismo , Encéfalo/metabolismo , Excitación Neurológica/metabolismo , Pentilenotetrazol/toxicidad , Convulsiones/metabolismo , Fases del Sueño/fisiología , Animales , Encéfalo/efectos de los fármacos , Excitación Neurológica/efectos de los fármacos , Masculino , Ratones , Ratones Noqueados , Convulsiones/inducido químicamente , Fases del Sueño/efectos de los fármacosRESUMEN
Interleukin-6-deficient (IL-6(-/-)) mice and their normal littermate (WT) were studied to evaluate their susceptibility to seizures induced by electroshock and audiogenic stimuli at different ages. No significant changes in maximal electroshock susceptibility were evidenced between the two strains, while audiogenic seizures (AGS) can be induced only in IL-6(-/-) mice. The effects of age and genetic condition on AGSs were evaluated. The behavioural and electrocortical changes during audiogenic stimulus were observed. In addition, the levels of neurotransmitter amino acids in five brain areas (of both strains) were measured at 60 days of age. Aspartate level significantly increased in the brain stem (BS) and hippocampus (HI), while it decreased in the diencephalon (DE) of IL-6(-/-) mice. Glutamate content significantly decreased in the cerebellum (CB), DE and HI. GABA levels significantly decreased in all the areas studied. Glycine significantly decreased in the BS, CB and DE, while taurine decreased only in the DE. The levels of glutamine significantly decreased in all the areas examined, except in the cortex (CX). The changes of neuroactive amino acid levels, particularly in the BS, might explain the characteristic of high propensity to AGS of IL-6(-/-) mice. The present data support the validity of IL-6(-/-) mice as a novel epileptic model for the study of the pathophysiology and pharmacology of epilepsy.
Asunto(s)
Estimulación Acústica/métodos , Encéfalo/metabolismo , Epilepsia Refleja/genética , Epilepsia Refleja/metabolismo , Predisposición Genética a la Enfermedad , Interleucina-6/deficiencia , Interleucina-6/genética , Neurotransmisores/biosíntesis , Factores de Edad , Animales , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Noqueados , Neurotransmisores/genéticaRESUMEN
Within the series of chiral 3,3'-(1,2-ethanediyl)bis[2-arylthiazolidin-4-ones], the 3,4-dimethoxyphenyl substituted derivative was found in the primary anti-inflammatory screening to be endowed with superior in vivo properties and good safety profile. Such a lead compound was modified by eliminating 3-methoxy group while retaining 4-methoxy group on the aryl rings at 2 and 2' stereogenic carbons. The 2R,2'S-meso isomer (VIG3b) of the resulting bisthiazolidinone has been widely investigated. The inhibitory effects on cyclo-oxygenase-1 and cyclo-oxygenase-2 isoenzymes were measured in a human whole blood assay. VIG3b was almost 50 times more selective on the inducible isoform. The cyclo-oxygenase-2 preferential selectivity has been confirmed by modeling VIG3b into the cyclo-oxygenase-1 and cyclo-oxygenase-2 active sites. Furthermore, VIG3b was assayed in the experimental model of carrageenan-induced lung injury by evaluating its ability to inhibit: (1) fluid accumulation in the pleural cavity, (2) neutrophil infiltration, (3) prostaglandin E(2) production and (4) lung injury. VIG3b exhibited interesting activity in all these tests.