RESUMEN
Despite the fact that a significant fraction of kidney graft dysfunctions observed after transplantation is due to ischemia-reperfusion injuries, there is still no clear consensus regarding optimal kidney preservation strategy. This stems directly from the fact that as of yet, the mechanisms underlying ischemia-reperfusion injury are poorly defined, and the role of each preservation parameter is not clearly outlined. In the meantime, as donor demography changes, organ quality is decreasing which directly increases the rate of poor outcome. This situation has an impact on clinical guidelines and impedes their possible harmonization in the transplant community, which has to move towards changing organ preservation paradigms: new concepts must emerge and the definition of a new range of adapted preservation method is of paramount importance. This review presents existing barriers in transplantation (e.g., temperature adjustment and adequate protocol, interest for oxygen addition during preservation, and clear procedure for organ perfusion during machine preservation), discusses the development of novel strategies to overcome them, and exposes the importance of identifying reliable biomarkers to monitor graft quality and predict short and long-term outcomes. Finally, perspectives in therapeutic strategies will also be presented, such as those based on stem cells and their derivatives and innovative models on which they would need to be properly tested.
Asunto(s)
Trasplante de Riñón , Riñón , Preservación de Órganos/métodos , Perfusión/métodos , Daño por Reperfusión/prevención & control , Animales , Humanos , Preservación de Órganos/efectos adversos , Perfusión/efectos adversos , Guías de Práctica Clínica como Asunto , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patologíaRESUMEN
The migration kinetics of bisphenol-A-diglycidyl-ether (BADGE) from processed and non-processed model cans into vegetable oil was investigated as a function of the process treatment and the temperature of storage. Cans were either not heat-treated at all or were processed at 115 degrees C for 30 min or for 1 h after filling with oil. Each series of experiments comprised 30 samples and was further divided into three groups to be stored at different temperatures (20, 40 and 60 degrees C). Aliquots from the samples were taken at regular intervals for > 1 year. Samples were analysed for BADGE by high-performance liquid chromatography with fluorescence detection. The results showed that temperature processing had the largest effect on migration of BADGE. Storage temperature also significantly influenced migration from non-processed cans, in particular at higher storage temperatures such as 60 degrees C. Some samples were Subjected to 60 degrees C storage after an initial period at 20 degrees C and an effect on migration was also noted, although to a much lesser extent than from processing. The results of migration at higher temperatures were also correlated to the potential degradation of BADGE from oxidation products.
Asunto(s)
Carcinógenos/química , Compuestos Epoxi/química , Contaminación de Alimentos/análisis , Embalaje de Alimentos , Compuestos de Bencidrilo , Manipulación de Alimentos/métodos , Conservación de Alimentos/métodos , Calor , Humanos , Aceites de Plantas/química , Temperatura , Factores de TiempoRESUMEN
It is generally assumed that bumetanide possesses some selectivity for the renal Na-K-Cl cotransporter NKCC2, although the results are scarce in the literature and comparisons were done with extra-renal NKCC1 at its basal, almost silent state. Here we investigated NKCC2/NKCC1 selectivity of loop diuretic drugs (bumetanide, piretanide and furosemide) as a function of the NKCC1 activated state (NKCC1 was activated by hypertonic media). NKCC2 activity was measured in isolated rat medullary thick ascending limb (mTAL) and NKCC1 in rat thymocytes and erythrocytes. When NKCC2 was compared with NKCC1at its activated state, all three diuretic drugs inhibited NKCC2 and NKCC1 with the same potency (bumetanide pIC50=6.48, 6.48 and 6.47; piretanide pIC50=5.97, 5.99 and 6.29; and furosemide pIC50=5.15, 5.04 and 5.21 for mTAL NKCC2, erythrocyte NKCC1 and thymocyte NKCC1, respectively). Basal NKCC1 exhibited a lower diuretic sensitivity, although with marked differences depending on the diuretic drug and the cell type in consideration and with the notable exception of furosemide in erythrocytes. Molecular modelling showed that bumetanide and piretanide possess four potentially active groups, of which three are shared with furosemide at similar intergroup distances. Of these three common groups, one should not bind to basal NKCC1 in thymocytes. The fourth (phenoxy) group (absent in furosemide) confers higher lipophilicity and should not bind to basal NKCC1 in erythrocytes. In conclusion, loop diuretics had no NKCC2/NKCC1 selectivity, when NKCC1 is measured at its activated state. Basal NKCC1 has a reduced diuretic sensitivity, of very different magnitude depending on the diuretic drug and cell type in consideration.
Asunto(s)
Diuréticos/farmacología , Transporte Iónico/efectos de los fármacos , Asa de la Nefrona/efectos de los fármacos , Simportadores de Cloruro de Sodio-Potasio/metabolismo , Animales , Bumetanida/química , Bumetanida/farmacología , Diuréticos/química , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Furosemida/química , Furosemida/farmacología , Asa de la Nefrona/metabolismo , Masculino , Ratas , Ratas Wistar , Miembro 1 de la Familia de Transportadores de Soluto 12 , Miembro 2 de la Familia de Transportadores de Soluto 12 , Sulfonamidas/química , Sulfonamidas/farmacología , Timo/citología , Timo/efectos de los fármacos , Timo/metabolismoRESUMEN
Calcium dobesilate possesses antioxidant properties and protects against capillary permeability by reactive oxygen species in the rat peritoneal cavity, but whether a similar action can take place in the diabetic rat retina is unknown. We investigated the oral treatment of diabetic rats with calcium dobesilate on the prevention of free radical-mediated retinal injury induced by ischemia/reperfusion (90 min ischemia followed by 3 min and/or 24 h of reperfusion). Streptozotocin-induced diabetic rats were orally treated with 50 and 100 mg/kg of calcium dobesilate for 10 days (n=12 in each group). In the first series of studies, calcium dobesilate was found to significantly reduce the maldistribution of ion content in diabetic ischemic/reperfused rat retina. Thus, in diabetic rats treated with 100 mg/kg/day calcium dobesilate, ischemia/reperfusion provoked: (i) 27.5% increase in retinal Na(+) content compared to 51.8% in the vehicle-treated group (P<0.05), and (ii) 59.6% increase in retinal Ca(2+) content compared to 107.1% in vehicle-treated animals (P<0.05). In the second series of studies, calcium dobesilate was found to significantly protect diabetic rat retina against inhibition of Na(+)/K(+)-ATPase and Ca(2+)/Mg(2+)-ATPase activities by ischemia/reperfusion (54% and 41% reduction, respectively, with 100 mg/kg of calcium dobesilate) and also against changes in retinal ATP, reduced glutathione (GSH), and oxidized glutathione (GSSG) contents. In the third series of experiments, rats treated with 100 mg/kg of calcium dobesilate reduced the hydroxyl radical signal intensity to 41% (measured by electron paramagnetic resonance), induced by ischemia/reperfusion in diabetic rat retina. Finally, 100 mg/kg calcium dobesilate significantly reduced retinal edema (measured by the thickness of the inner plexiform layer) in diabetic rats. In conclusion, oral treatment with calcium dobesilate significantly protected diabetic rat retina against oxidative stress induced by ischemia/reperfusion. Whether the antioxidant properties of calcium dobesilate explain, at least in part, its beneficial therapeutic effects in diabetic retinopathy deserves further investigation.
Asunto(s)
Antioxidantes/farmacología , Dobesilato de Calcio/farmacología , Diabetes Mellitus Experimental/complicaciones , Retinopatía Diabética/prevención & control , Daño por Reperfusión/complicaciones , Adenosina Trifosfato/metabolismo , Animales , Antioxidantes/uso terapéutico , ATPasa de Ca(2+) y Mg(2+)/efectos de los fármacos , ATPasa de Ca(2+) y Mg(2+)/metabolismo , Calcio/metabolismo , Dobesilato de Calcio/uso terapéutico , Cationes/metabolismo , Diabetes Mellitus Experimental/metabolismo , Retinopatía Diabética/etiología , Retinopatía Diabética/metabolismo , Radicales Libres/metabolismo , Glutatión/efectos de los fármacos , Glutatión/metabolismo , Disulfuro de Glutatión/efectos de los fármacos , Disulfuro de Glutatión/metabolismo , Magnesio/metabolismo , Masculino , Potasio/metabolismo , Ratas , Ratas Sprague-Dawley , Retina/efectos de los fármacos , Retina/metabolismo , Retina/patología , Sodio/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/efectos de los fármacos , ATPasa Intercambiadora de Sodio-Potasio/metabolismoRESUMEN
The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPLC with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1 mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether) analysed in the same products in the context of a previous study.
Asunto(s)
Carcinógenos/análisis , Compuestos Epoxi/análisis , Resinas Epoxi/análisis , Peces , Contaminación de Alimentos/análisis , Conservación de Alimentos , Animales , Compuestos de Bencidrilo , Carcinógenos/química , Cromatografía Líquida de Alta Presión , Compuestos Epoxi/química , Resinas Epoxi/química , Unión Europea , Embalaje de Alimentos , Isomerismo , Aceites , SuizaRESUMEN
A survey at the European levels was initiated on the quantification of bisphenol-A-diglycidyl-ether (BADGE) in canned fish in oil in order to assess the exposure of BADGE. A total of 382 canned fish sample were collected from all 15 Member States and Switzerland and analysed for BADGE in fish. The fish was extracted first with hexane and reextracted with acetonitrile, followed by a membrane filtration and reverse phase HPLC analysis with fluorescence detection. The analysis of the fish showed that about 3% of the samples contained BADGE at a level above 1 mg/kg. The samples exceeding the limit by a larger margin were mostly from anchovy cans and cans manufactured in 1991-1995.
Asunto(s)
Carcinógenos/análisis , Compuestos Epoxi/análisis , Peces , Contaminación de Alimentos/análisis , Conservación de Alimentos , Animales , Compuestos de Bencidrilo , Cromatografía Líquida de Alta Presión , Unión Europea , Humanos , AceitesRESUMEN
Within the framework of the AIR3-CT94-2360 EU-project, the stability of three plastics additives in three EU aqueous and fatty food simulants and in two alternative simulants was studied under various time-temperature conditions. The additives tested were bis(2-ethylhexyl) adipate (DEHA), bis(2-ethylhexyl) phthalate (DEHP) and octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propionate (Irganox 1076). The various test conditions included exposures of 10 days at 40 degrees C, 1 h at reflux temperature for all aqueous simulants, 10 days at 40 degrees C and 1 h 175 degrees C for the olive oil and 2 days at 20 degrees C and 3 h at 60 degrees C for the isooctane simulant. Following the exposure, the additive samples were extracted from aqueous simulants with hexane. A sonication step was necessary to ensure maximum extraction of control samples. In the case of the isooctane simulant, the samples were analyzed directly from the simulant. The oil samples were extracted by acetonitrile. The extracts of samples exposed to various heat conditions as well as unexposed spiked controls and blanks were analysed by gas chromatography (GC) on a non-polar (5%--phenyl)-methylpolysiloxane capillary column with high temperature capabilities. The results showed that DEHA, DEHP and Irganox 1076 were stable at 40 degrees C and at reflux temperature in ethanolic or acidic aqueous simulants. The various additives were also stable in the organic isooctane simulant as well as in the fatty simulant olive oil. Studies on the stability of such additives used in food packaging are designed for regulatory purposes as an aid to decide whether the legislation should regulate limits for plasticizers based on a quantity in the food packaging itself or based on an ingested dose by the consumer.
Asunto(s)
Contaminación de Alimentos , Embalaje de Alimentos , Plastificantes , Cromatografía de Gases , Calor , Modelos QuímicosRESUMEN
In order to delineate ion transport mechanisms involved in volume homeostasis of freshly isolated newborn rat ventricular myocytes, we investigated the effects of ion substitutions and pharmacological maneuvers upon (1) isotonic volume, (2) hypotonically induced initial swelling, and (3) the subsequent regulatory volume decrease (RVD), as determined by electronic cell sizing. Cardiomyocytes exposed to hypotonic medium (176 mosmol/l) swelled by 51+/-1% of isotonic volume, and they underwent a partial regulatory volume decrease (RVD), reaching a maximum regulation after 30 min (51+/-1% of initial swelling), with a half-time (t1/2) of 6+/-1 min (n=60). RVD was associated with significant cardiomyocyte K+ loss (12+/-4% at 5 min and 15+/-2% of isotonic control after 30 min: n=6, P<0.001), 71% of which was Cl- dependent (P<0.05). Within the 30-min experimental time frame, ouabain, a Na+/K+ pump inhibitor, had no significant effect on RVD (despite an inhibitory trend), cell swelling or on isotonic volume (n=6). Bumetanide (50 microM), a Na+-K+-Cl- co-transport blocker, induced a significant reduction of isotonic cell volume (3+/-2%, n=6. P<0.05), potentiated initial swelling by 16+/-1% (n=8, P<0.02), and it partially inhibited RVD (24+/-11% at 30 min, n=6), whereas Na+ omission had no significant effect on isotonic cell volume, cell swelling or RVD. The effects of bumetanide on initial swelling and RVD were prevented by gadolinium ion (10 microM), a stretch-activated cation channel blocker (n=5). Quinidine (500 microM), a non-selective Ca(2+)-activated potassium channel blocker with no side-effects on K(+)-Cl(-) cotransport, did not modify initial cell swelling, but inhibited RVD (50+/-3% at 5 min, n=9, P<0.01; 22+/-3% at 30 min), an effect which was cancelled by external Ca2+ chelation with EGTA (n=5), and reproduced by tetraethylammonium (TEA, 20 mM), another K+ channel blocker. 4,4'-Diisothiocyanatostilbene 2,2'-disulfonic acid (DIDS, 100 microM), a non-selective swelling-activated Cl- channel blocker with marginal side-effects on K(+)-Cl(-)cotransport, did not modify initial swelling, but inhibited RVD to the same extent as quinidine (42+/-3% at 5 min, and 23+/-3% at 30 min, n=15, P<0.05), whereas hypotonic Cl(-)-free solution had no effect on isotonic volume, but potentiated initial swelling by 16+/-2% (P<0.05) and fully inhibited RVD (n=5, P<0.001). R(+)-[(2-n-Butyl-6,7-dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1H-inde n-5yl)-oxy] acetic acid) (DIOA, 80 microM), a K(+)-Cl- cotransport blocker (with inhibitory potency toward Ca(2+)-activated K+ channels), inhibited 87+/-5% of the RVD process at 5 min (P<0.001) and 56+/-16% at 30 min (P<0.001), whereas it had a small effect on isotonic volume (+4%, P<0.01) and initial cell swelling (+2%, N.S.; n=9). In contrast to quinidine, DIOA was able to inhibit Ca(2+)-omission-resistant RVD (full inhibition at 5 min, and 56+/-9% at 30 min; P<0.01, n=5). In conclusion, our results suggest that at least three distinct ion transport mechanisms are involved in the RVD in newborn rat cardiomyocytes: (1) K+ and Cl-channels, (2) K(+)-Cl- cotransport, and (3) Na(+)-K(+)-Cl- co-transport.
Asunto(s)
Animales Recién Nacidos/fisiología , Proteínas Portadoras/metabolismo , Miocardio/citología , Miocardio/metabolismo , Canales de Potasio/metabolismo , Simportadores , Animales , Proteínas Portadoras/antagonistas & inhibidores , Tamaño de la Célula/fisiología , Canales de Cloruro/antagonistas & inhibidores , Canales de Cloruro/metabolismo , Cloruros/metabolismo , Medios de Cultivo , Corazón/efectos de los fármacos , Soluciones Hipotónicas , Técnicas In Vitro , Cinética , Potasio/metabolismo , Bloqueadores de los Canales de Potasio , Ratas , Ratas Wistar , ATPasa Intercambiadora de Sodio-Potasio/antagonistas & inhibidores , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Cotransportadores de K ClRESUMEN
Calcium dobesilate was tested in rats with streptozotocin(STZ)-induced diabetes, for its angioprotective properties against the extravasation of plasma Evans blue towards the peritoneal cavity, induced in situ by the free radical generating agent phenazine methosulfate (PMS). In diabetic rats pretreated with a single oral dose of 50 and 200 mg/kg calcium dobesilate, PMS-induced Evans blue extravasation was respectively reduced by 60 and 100% with respect to values in vehicle-treated animals (p < 0.05 and p < 0.01 respectively; in diabetic rats, PMS-induced Evans blue peritoneal extravasation was double as in control animals = 0.0814 +/- 0.019 vs. 0.0396 +/- 0.0083 h-1, p < 0.05). In diabetic rats pretreated with 50 and 100 mg/kg/day of calcium dobesilate for 7 days, PMS-induced Evans blue extravasation was respectively reduced by 56 and 80% with respect to values in vehicle-treated animals (p < 0.01 and p < 0.001 respectively). In conclusion, calcium dobesilate p.o. significantly and dose-dependently antagonized the increase in capillary permeability induced by an oxidative stress in the peritoneal cavity of diabetic rats. These results further suggest that the antioxidant properties of calcium dobesilate can be involved, at least in part, in its angioprotective actions in humans.
RESUMEN
Calcium dobesilate possesses antioxidant properties in vitro, but the in vivo significance and putative angioprotective role of these properties are undefined. Here, calcium dobesilate was tested in a newly developed in vivo model of microvascular permeabilization induced by reactive oxygen species in the rat peritoneal cavity. In this model, microvascular permeabilization is equated to the rate of Evans blue extravasation toward the peritoneal cavity. Basal Evans blue extravasation (rate constant values ke = 0.0176 +/- 0.0015 h-1) was markedly and significantly increased by reactive oxygen species generated in situ, with: (i) phenazine methosulfate/NADH (delta ke(phenazine methosulfate) = 0.0419 +/- 0.0043 h-1) and (ii) xanthine/xanthine oxidase (delta ke(xo) = 0.0383 +/- 0.0010x h-1). These actions of reactive oxygen species were abolished by locally injected superoxide dismutase (i.p., 300 units/kg). Intraperitoneally given calcium dobesilate (100 mg/kg) inhibited 75-100% of reactive oxygen species-induced Evans blue extravasation. By the intravenous route, calcium dobesilate i.v. (1-50 mg/kg) dose dependently inhibited phenazine methosulfate-induced Evans blue extravasation with an ID50 of 2-5 mg/kg (full inhibition was reached at 20-50 mg/kg). After single oral administration, calcium dobesilate (5-500 mg/kg) dose dependently inhibited phenazine methosulfate-dependent Evans blue extravasation with an ID50 of 50-100 mg/kg (81% inhibition at 500 mg/kg, P < 0.003). After 7 days of oral calcium dobesilate (50 mg/kg once/day) phenazine methosulfate-induced Evans blue peritoneal extravasation was significantly reduced by half. These effects of calcium dobesilate were similar to those observed with a comparative antioxidant molecule, rutin. In conclusion, rat peritoneal microvascular permeability was strongly increased by reactive oxygen species, an effect that was significantly reduced by intraperitoneal, intravenous and oral calcium dobesilate. These results support the hypothesis that the antioxidant properties of calcium dobesilate could play a role in its angioprotective properties in vivo.
Asunto(s)
Dobesilato de Calcio/farmacología , Permeabilidad Capilar/efectos de los fármacos , Hemostáticos/farmacología , Especies Reactivas de Oxígeno/fisiología , Administración Oral , Animales , Dobesilato de Calcio/química , Dobesilato de Calcio/uso terapéutico , Permeabilidad Capilar/fisiología , Relación Dosis-Respuesta a Droga , Vías de Administración de Medicamentos , Azul de Evans , Extravasación de Materiales Terapéuticos y Diagnósticos/fisiopatología , Extravasación de Materiales Terapéuticos y Diagnósticos/prevención & control , Hemostáticos/uso terapéutico , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Masculino , Cavidad Peritoneal/fisiopatología , Ratas , Ratas WistarRESUMEN
The effect of cell swelling on intracellular calcium concentration ([Ca2+]i) was studied in newborn rat cardiomyocytes. Hypotonic cell swelling induced a fast and transient [Ca2+]i increase (hypotonically induced calcium increase, HICI; 388±47 nM, n=14). HICI was not inhibited by cyclopiazonic acid (CPA), an inhibitor of sarcoplasmic Ca2+-ATPase, nor ryanodine (an inhibitor of calcium-induced calcium release), whereas it was abolished (11±19 nM, n=5) in the absence of external calcium. Thus, HICI appeared to depend exclusively on entry of external calcium. Gadolinium ion (Gd3+), a generic inhibitor of stretch-activated cation channels (SACs), was unable to affect HICI (353±79 nM, n=6). Similarly, HICI was unaffected by internal Na+ depletion and external Na+ omission. These results suggest that neither Gd3+-sensitive SACs nor Na+-Ca2+ exchange is responsible for HICI. Conversely, HICI was inhibited by diltiazem (42±4 nM, n=3) and by membrane predepolarization (40±18 nM, n=5), suggesting an involvement of L-type voltage-activated calciumchannels. Cardiomyocyte swelling was followed by a regulatory volume decrease (RVD). The putative role of HICI in volume regulation was studied by removal of external calcium. This procedure significantly slowed RVD but did not abolish it. In conclusion, newborn rat cardiomyocytes exhibit an external-calcium-dependent HICI which contributes partially to the RVD.
Asunto(s)
Calcio/metabolismo , Tamaño de la Célula , Soluciones Hipotónicas , Miocitos Cardíacos/citología , Miocitos Cardíacos/metabolismo , Animales , Animales Recién Nacidos , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio/metabolismo , Diltiazem/farmacología , Colorantes Fluorescentes/metabolismo , Fura-2/metabolismo , Gadolinio/metabolismo , Potenciales de la Membrana/fisiología , Miocitos Cardíacos/efectos de los fármacos , Concentración Osmolar , Ratas , Sodio/metabolismoRESUMEN
BACKGROUND: It has been proposed that ischemic coronary disease (ICD) associated potassium loss could be due to modifications of potassium permeability. We investigated whether a positive family history of ICD can influence this parameter. We have compared potassium permeability in erythrocytes from ICD patients and from positive family history subjects (FICD) with control subjects. METHODS: All patients and subjects were carefully selected for the absence of hypertension and dysmetabolic pathologies. ICD group: 24 patients (19 males, 5 females; ages 43 to 69) all affected by ischemic coronary disease, under no drug treatment; FICD group: 18 subjects (all males, ages 27 to 42) with a verified positive ICD family history, without hypertensive family history and cardiovascular pathology; control group: 16 subjects (11 males, 5 females; ages 28 to 48) without positive family history of ICD. Passive potassium efflux (PPE) was spectrophotometrically measured in K-free medium containing ouabain and bumetanide. The kinetic constant was calculated by dividing PPE by the erythrocyte potassium concentration. RESULTS: No statistically significant differences were noted between the intracellular potassium content of the three groups. However, (1) the passive potassium permeability of the ICD group was significantly higher (kK=0.055 +/- 0.021 h(-1), n=24) than that of the control group (kK=0.023 +/- 0.008 h(-1), n= 16; p<0.00001), (2) the FICD group was higher (kK=0.036 +/- 0.012 h(-1), n=18) than the control group (p<0.001), and (3) the ICD group was higher than the FICD group (p<0.001). CONCLUSIONS: Our results suggest an inheritability of ICD, paralleling the familial aggregation of the pathology. Erythrocyte potassium permeability could represent an early marker of ischemic coronary disease and be used as a prophylactic tool.
Asunto(s)
Eritrocitos/metabolismo , Isquemia Miocárdica/metabolismo , Potasio/metabolismo , Adulto , Anciano , Bumetanida/farmacología , Permeabilidad de la Membrana Celular , Membrana Eritrocítica/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/diagnóstico , Ouabaína/farmacología , ATPasa Intercambiadora de Sodio-Potasio/efectos de los fármacos , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Espectrofotometría AtómicaRESUMEN
Calcium dobesilate, a vascular protective agent, was tested in vitro for its scavenging action against oxygen free radicals. Calcium dobesilate was as potent as rutin to scavenge hydroxyl radicals (IC50 = 1.1 vs 0.7 microM, respectively). It was also able to scavenge superoxide radicals, but with 23 times less potency than rutin (IC50 = 682 vs 30 microM, respectively). Calcium dobesilate significantly reduced platelet activating factor (PAF)-induced chemiluminescence in human PMN cells and lipid peroxidation by oxygen free radicals in human erythrocyte membranes, although these actions required calcium dobesilate concentrations > or = 50 microM. Finally, in cultured bovine aortic endothelial cells, magnesium dobesilate reduced the increase in cytosolic free calcium induced by hydrogen peroxide and inhibited phenazine methosulfate-induced cell potassium loss. In conclusion, calcium dobesilate was effective in scavenging hydroxyl radicals in vitro, at therapeutically relevant concentrations. Conversely, higher concentrations of the compound were required to scavenge superoxide radicals or to protect the cells against the deleterious effects of intracellular reactive oxygen species. Further studies in vivo are required to determine if these antioxidant properties of calcium dobesilate can play a role in its vascular protective mechanisms.
Asunto(s)
Antioxidantes/farmacología , Dobesilato de Calcio/farmacología , Depuradores de Radicales Libres/farmacología , Animales , Aorta/efectos de los fármacos , Calcio/metabolismo , Bovinos , Células Cultivadas , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Membrana Eritrocítica/efectos de los fármacos , Hemostáticos/farmacología , Humanos , Radical Hidroxilo/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Mediciones Luminiscentes , Neutrófilos/efectos de los fármacos , Factor de Activación Plaquetaria , Potasio/metabolismo , Superóxidos/metabolismo , Xantina/análisis , Xantina Oxidasa/análisisRESUMEN
Fenspiride inhibits the calcium signal evoked by the inflammatory peptide formyl-Met-Leu-Phe (fMLP) in peritoneal macrophages, but at concentrations (approximately 1 mM) far above the therapeutic range (approximately 1 microM). Here, in rat alveolar macrophages, high fenspiride concentrations (1 mM) were required to inhibit the calcium signals evoked by the calcium agonist Bay K8644 or by ionomycin. Moreover, fenspiride (1 mM) was a poor inhibitor of the cell membrane depolarization induced by gramicidine D. By contrast, fenspiride blocked Na+-H+ antiport activation by (i) fMLP with an IC50 = 3.1 +/- 1.9 nM and (ii) PMA (phorbol 12-myristate 13-acetate) with an IC50 = 9.2 +/- 3.1 nM. Finally, protein kinase C (PKC) activity of macrophage homogenate was not significantly modified by 10 or 100 microM fenspiride (at 100 microM: 2.57 +/- 1.60 vs. 2.80 +/- 1.71 pmol/10(6) cells/min). In conclusion, fenspiride inhibits fMLP- and PMA-induced pH signals in rat alveolar macrophages, probably by acting distally on the PKC transduction signal. This pH antagonistic action may be relevant for the antiinflammatory mechanism of fenspiride and requires further investigation.
Asunto(s)
Broncodilatadores/farmacología , Macrófagos Alveolares/efectos de los fármacos , Compuestos de Espiro/farmacología , Animales , Calcio/metabolismo , Membrana Celular/efectos de los fármacos , Concentración de Iones de Hidrógeno , N-Formilmetionina Leucil-Fenilalanina , Ratas , Transducción de Señal/efectos de los fármacos , Acetato de TetradecanoilforbolRESUMEN
The diuretic drug xipamide improves myocardial relaxation in hypertensive patients with left ventricular hypertrophy, but its mechanism of action is unknown. Here, xipamide was tested in cultured rat heart myogenic H9c2 cells and newborn cardiomyocytes for its effects on cell acidification (and Ca2+ mobilization). In H9c2 cells, blocking Na+/H+ exchange with amiloride (2 mM) provoked cell acidification with rate = 0.82 +/- 0.17 pH units/h (n = 6). Xipamide 1 microM maximally inhibited 50 +/- 7% (n = 9) of cell acidification. The action of xipamide required the presence of HCO3- and was antagonized by the HCO3(-)-transport blocker DIDS (4,4'-diisothiocyanostilbene-2.2'-disulfonic acid). Conversely, the carbonic anhydrase (EC 4.2.1.1) inhibitor acetazolamide failed to prevent xipamide action. Finally, xipamide was without significant effect on the Ca2+ signals induced by endothelin-1, vasopressin or the Ca2+ ionophore ionomycin. In newborn rat cardiomyocytes, xipamide reduced amiloride-induced cell acidification at similar concentrations as in H9c2 cardiocytes, but with a slightly higher extent of maximal inhibition (70-80%). In conclusion, xipamide reduced amiloride-dependent cell acidification in the rat heart myogenic H9c2 cell line and in newborn rat cultured cardiomyocytes. This action of xipamide seems to be related to a complex interaction with DIDS-sensitive HCO3- movements. Prevention of cell acidification by xipamide could be involved in the beneficial effects of this compound in myocardial relaxation and left ventricle filling in hypertensive patients with left ventricular hypertrophy.
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Amilorida/antagonistas & inhibidores , Diuréticos/antagonistas & inhibidores , Diuréticos/farmacología , Corazón/efectos de los fármacos , Xipamida/farmacología , Ácido 4,4'-Diisotiocianostilbeno-2,2'-Disulfónico/farmacología , Animales , Calcio/metabolismo , Anhidrasas Carbónicas/efectos de los fármacos , Anhidrasas Carbónicas/metabolismo , Células Cultivadas/efectos de los fármacos , Concentración de Iones de Hidrógeno , Miocardio/citología , Miocardio/metabolismo , RatasRESUMEN
We have recently described that urines from salt-loaded rats contain a potent natriuretic factor acting at the Na-K-Cl cotransport system (CIF: "Cotransport Inhibitory Factor"). Here we investigated the kinetics of the urinary CIF excretion which follows an oral salt-load: (i) in normal rats, relative to that of the atrial natriuretic peptide (ANP) and (ii) in an experimental model of salt-dependent genetic hypertension (Dahl's rats). Thus, Wistar rats were orally loaded with 2% NaCl for 8 days. Urinary CIF excretion was measured by testing the inhibitory potency of urines on bumetanide-sensitive lithium efflux in lithium-loaded human erythrocytes. Plasmatic levels of ANP were measured by radioimmunoassay. Plasma ANP rapidly and transiently increased during the first 24 hs of salt-load, decreasing thereafter down to normal levels in 6-8 days. Conversely, CIF slowly increased after 24 hs up to maximal constant levels after 5 days of salt-loading. Dahl salt-sensitive rats exhibited highly significant increases in urinary CIF excretion with respect to salt-resistant rats. In the basal state (before salt-loading) urinary CIF excretion was 101 +/- 13 vs 17.6 +/- 4.5 units/day in salt-sensitive vs. salt-resistant rats (n = 7 for each group, p < 0.001). This difference was maintained after salt loading (3 380 +/- 990 vs. 456 +/- 159 units/day, p < 0.05 at day 5).(ABSTRACT TRUNCATED AT 250 WORDS)
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Factor Natriurético Atrial/farmacocinética , Animales , Factor Natriurético Atrial/fisiología , Hipertensión/metabolismo , Asa de la Nefrona/metabolismo , Masculino , Natriuresis , Ratas , Ratas WistarRESUMEN
INTRODUCTION: Clinical studies suggest that lacidipine (LA) is better tolerated than other DHP, in terms of peripheral edema. We evaluated edema due to LA, nitrendipine (NT) and nifedipine (NF) in SHR. METHODS: Mean arterial pressure (MAP) was measured with an intra-femoral probe. Peripheral edema was determined (i) by the plasmatic distribution of 14C-albumin, (ii) by Evans blue extravasation. RESULTS: In bolus(ip), LA, NT and NF had non different effects on plasmatic *ALB, i.e. + 3.9 +/- 1.7 (delta % vs control at 60 min; mean +/- SEM, n = 18). Evans blue extravasation (hind paws muscle = EBM) was positively correlated to MAP reduction (EBM = 0.1 x delta MAP + 5.2; p < 0.025), without differences between the molecules. In chronical administration (9 days), at comparable MAP decreases (31 +/- 2 mmHg), there was less edema formation with LA (0.05 mg/kg/j) than with NT (0.5 mg/kg/j) or NF (1.4 mg/kg/j): the variations of *ALB were respectively (% vs control at 45 min after tracer injection; mean +/- SD): + 5% (n = 10) vs. 73% (n = 14; p < 0.01 vs LA) and + 34% (n = 10; p < 0.01 vs LA); no significant change of hematocrit or plasma volume was noted. CONCLUSION: Our results confirm, in SHR, that lacidipine induces a very moderate edema formation. This does not seem to be due to a renal effect, nor to an effect on peripheral resistances. It was only observed in chronical administration, which suggests that pharmacokinetic properties of lacidipine are involved.
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Bloqueadores de los Canales de Calcio/efectos adversos , Dihidropiridinas/efectos adversos , Edema/inducido químicamente , Nifedipino/efectos adversos , Nitrendipino/efectos adversos , Animales , Presión Sanguínea/efectos de los fármacos , Dihidropiridinas/farmacología , Nifedipino/farmacología , Nitrendipino/farmacología , Ratas , Ratas Endogámicas SHR , Albúmina Sérica/análisisAsunto(s)
Cloruros/sangre , Natriuréticos/orina , Potasio/sangre , Cloruro de Sodio Dietético/administración & dosificación , Sodio/sangre , Animales , Eritrocitos/metabolismo , Hipertensión/sangre , Hipertensión/etiología , Hipertensión/orina , Transporte Iónico/fisiología , Masculino , Ratas , Ratas WistarRESUMEN
1. Hypotonic stress unmasked inward and outward K+ and Cl- movements in rat thymocytes. This KCl flux stimulation was reduced by DIOA (dihydroindenyl-oxy-alkanoic acid), but not by DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonate), quinidine, DPAC 144 (5-nitro-2-(2-phenylethyl-amino)-benzoic acid), bumetanide or ouabain. 2. In isotonic media (308 +/- 5 mosmol kg-1), the cells exhibited the following DIOA-sensitive fluxes: (i) a K+ efflux of 42.7 +/- 17.1 mmol (l cells.h)-1 (mean +/- S.D., n = 7), (ii) a Cl- efflux of 68 +/- 21 mmol (l cells.h)-1 (n = 3), (iii) a Rb+ influx of 9.7 +/- 3.9 mmol (l cells.h)-1 (n = 6) and (iv) a Cl- influx of 9.4 +/- 4.1 mmol (l cells.h)-1 (n = 6). 3. Hypotonic shock (183-200 mosmol kg-1) induced a sevenfold stimulation of DIOA-sensitive K+ and Cl- effluxes and a twofold stimulation of DIOA-sensitive Rb+ and Cl- influxes (with a Rb+ to Cl- stoichiometry of 1.04 +/- 0.31; mean +/- S.D., n = 6). 4. The DIOA-sensitive membrane carrier catalysed net outward KCl extrusion (the outward/inward flux ratio was 5-7 in isotonic media and 20 in hypotonic media at 189 mosmol kg-1). Inhibition of DIOA-sensitive 36Cl- efflux by cell K+ depletion suggested coupling of outward K+ and Cl- fluxes. Conversely, inward K+ and Cl- fluxes were found to be uncoupled in NO3- media and in K(+)-free media. 5. The results clearly show that rat thymocyte membranes possess a 1:1 K(+)-Cl- co-transport system which is strongly activated by hypotonic shock and catalyses net KCl extrusion.
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Cloruros/metabolismo , Potasio/metabolismo , Timo/metabolismo , Animales , Calcio/metabolismo , Ácidos Carboxílicos/farmacología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Cloro , Citosol/metabolismo , Concentración de Iones de Hidrógeno , Soluciones Hipotónicas , Técnicas In Vitro , Indenos/farmacología , Masculino , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Canales de Potasio/efectos de los fármacos , Canales de Potasio/metabolismo , Radioisótopos , Ratas , Ratas Wistar , Rubidio/metabolismo , Timo/citología , Timo/efectos de los fármacosRESUMEN
1. DIOA (dihydroindenyl-oxy-alkanoic acid), a potent inhibitor of the K(+)-Cl- co-transport system, fully blocked regulatory volume decrease (RVD) in swelled rat thymocytes, with an IC50 of 2.2 +/- 0.5 x 10(-5) mol l-1 (mean +/- S.D., n = 4). Conversely, RVD was resistant to quinine, quinidine, apamin, cetiedil, amiloride, bumetanide and DIDS (4,4'-diisothiocyanostilbene-2,2'-disulphonate). 2. DIOA-sensitive RVD followed mono-exponential kinetics, with t1/2 (half-lifetime) of 1-3 min and maximal capacity (Cmax) of about 55% of the initial cell swelling. Cmax and the initial rate of RVD (Vo) were both linear functions of the increase in cell volume. 3. RVD was: (i) slightly increased by replacing external Cl- by NO3-, (ii) reversed by replacing external Na+ by K+ (in the presence of external Cl-) and (iii) inhibited by cell K+ depletion. All these phenomena were blocked by DIOA (86 mumol l-1). 4. Increased membrane potassium permeability by valinomycin was unable to accelerate RVD or RVD reversal. 5. In the presence of DIOA, thymocytes responded like osmometers (the relative cell volume was a linear function of the reciprocal of the relative osmolality) in a large range of osmolalities. 6. The results strongly suggest that RVD in rat thymocytes is mediated by the K(+)-Cl- co-transport system.