RESUMEN
During exercise, the plasma urate levels and urinary excretion increase due to the enhanced purine degradation in skeletal muscle. Although urate transporter-1 (URAT1) is the main transporter responsible for the reabsorption of filtered urate, potential changes in its activity and expression during exercise have not been studied yet. Therefore, the effect of heavy muscle activity on renal URAT1 activity and expression was investigated in this study. Wistar rats were used in the study and the experimental design consisted of three groups: a control group, an exercise group where animals were exhausted once a day for 5 days, and a hyperuricemia group, which was induced by an uricase inhibitor, oxonic acid. URAT1 activity measurements were performed in isolated proximal tubule segments and expression of URAT1 mRNA and protein levels were determined by the reverse transcription polymerase chain reaction and western blot analyses, respectively. Increased citrate synthase activity in soleus muscle of exercised animals proved the efficiency of our exercise protocol. Proteinuria, glucosuria, and hypoglycemia were observed only in exercised animals; however, plasma and urinary urate levels were found to be elevated in both exercising and hyperuricemia groups. Moreover, in both of the groups URAT1 transporter activity was found to be increased despite the significant decrease in URAT1 protein levels. Considering the similar changes of urate metabolism observed in both exercising and hyperuricemic rats, our results suggest that exercise-induced changes in URAT1 expression and activity depend on the increased urate concentration in plasma.
Asunto(s)
Proteínas de Transporte de Anión/fisiología , Riñón/metabolismo , Esfuerzo Físico/fisiología , Animales , Proteínas de Transporte de Anión/biosíntesis , Masculino , Ratas , Ratas WistarRESUMEN
Cytoresistance is the term used to describe the response of the proximal tubule cells to various stress inducers via cholesterol accumulation. However, the role of extensive exercise as a renal insult has not been examined. In this study, the effect of heavy muscle activity on proximal tubule cytoresistance was investigated. Results obtained from rats subjected to running a treadmill for five days were compared to those of controls. Extensive muscle activity-induced soleus citrate synthase and blood lactate elevation were associated with normal MAP, RBF, and GFR. Blood electrolytes and cholesterol levels remained unchanged, whereas the total and free cholesterol accumulations in the proximal tubule cells of the exercised group were higher than controls. Cholesterol-loaded tubules were more resistant (as proved by LDH release) to an ATP-depleted/calcium overloaded second stress. These data clearly demonstrate that heavy muscle activity induces cholesterol accumulation in the proximal tubules of kidney, without influencing ATP generation.
Asunto(s)
Adenosina Trifosfato/metabolismo , Colesterol/metabolismo , Túbulos Renales Proximales/metabolismo , Túbulos Renales Proximales/patología , Carrera/fisiología , Animales , Modelos Animales de Enfermedad , Riñón/metabolismo , Masculino , Músculo Esquelético/metabolismo , Ratas , Ratas WistarRESUMEN
When the body is exposed to insults, the kidneys exhibit adaptive changes termed renal cytoresistance, characterized by cholesterol accumulation in the membranes of the tubule cells. However, heavy muscle activity has not yet been accepted as one of the stressors that could lead to cytoresistance. In order to study the renal functional characteristics of animals exposed to heavy muscle activity, rats were subjected to exhaustive treadmill exercise for 5 days and their data was compared to those of sedentary controls. It was found that in exercised rats, blood lactate, muscle citrate synthase and proximal tubule peroxynitrite levels were all elevated, suggesting the presence of oxidative stress in the proximal tubule segments. However, mean arterial pressure, renal blood flow, glomerular filtration rate, fractional excretion of sodium and potassium, and organic anion excretion remained normal. Despite unchanged blood cholesterol levels, cholesterol loading in the proximal tubule segments, especially the free form, and decreased lactate dehydrogenase release from cytoresistant proximal tubule segments indicated the development of renal cytoresistance. However, this resistance did not seem to have protected the kidneys as expected because organic anion accumulation associated with glycosuria and proteinuria, in addition to the elevated urinary cholesterol levels, all imply the presence of an impaired glomerular permeability and reabsorption in the proximal tubule cells. Therefore, we suggest that in response to heavy muscle activity the tubular secretion may remain intact, although cytoresistance in the proximal tubule cells may affect the tubular reabsorptive functions and basolateral uptake of substances. Thus, this differential sensitivity in the cytoresistance should be taken into account during functional evaluation of the kidneys. Key pointsThe cholesterol loading and decreased LDH release from PTSs isolated from exhausted rats indicate the heavy muscle activity induced renal cytoresistance.Heavy muscle activity-induced renal cytoresistance did not preserve the kidney functions.Organic anion accumulation as well as failure in the absorptive capacity of the tubule cells suggest the presence of some biochemical changes and elevated vulnerability of kidneys against nephrotoxic agents in rats subjected to heavy muscle activity.
RESUMEN
Considering the importance of nitric oxide generation in the regulation of vessel tone, reduced endothelial nitric oxide synthase (eNOS) expression in alveolar macrophages exposed to short-term silica (Si) suggests the possibility of Si-induced changes in endothelial functions. In this experimental study, the functional changes of the endothelial cells were investigated in the aortic rings of rats subjected to 50 mg Si/kg body weight in their drinking water for 8 days. Norepinephrine elicited contractility and dilation response to acetylcholine (ACh) was significantly high in the aortic rings of Si-treated group. Alteration in receptor-independent endothelial response to A23187 in the aortic rings of Si-exposed rats was less obvious, but sodium nitroprusside (SNP)-elicited dilation was reduced significantly. A23187-induced relaxation was fully eliminated with N-nitro-L-arginine methyl ester (L-NAME) pretreatment, whereas 19.24 +/- 4.36% of ACh response was L-NAME resistant and eliminated with 10-5 M tetraethylammonium (TEA). Despite a significant reduction in the share of NO, the contribution of indomethacine (IND)-sensitive relaxation to ACh response remained unchanged in Si group. As a result, our findings demonstrated that Si both modifies the characteristics of endothelial relaxants and attenuates smooth muscle cell responsiveness to NO. Si-induced reduced NO association with elevated endothelium-derived hyperpolarizing factor (EDHF) in response to ACh, together with reduced NO sensitization, might have clinical importance in cardiovascular pathology.
Asunto(s)
Aorta/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , Óxido Nítrico/metabolismo , Dióxido de Silicio/farmacología , Vasodilatación/efectos de los fármacos , Animales , Antiinflamatorios no Esteroideos/farmacología , Aorta/metabolismo , Aorta/fisiopatología , Factores Biológicos/metabolismo , Suplementos Dietéticos , Interacciones Farmacológicas , Células Endoteliales/metabolismo , Inhibidores Enzimáticos/farmacología , Alimentos Formulados , Masculino , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/fisiopatología , NG-Nitroarginina Metil Éster/farmacología , Donantes de Óxido Nítrico/farmacología , Técnicas de Cultivo de Órganos , Bloqueadores de los Canales de Potasio/farmacología , Ratas , Dióxido de Silicio/administración & dosificación , Tetraetilamonio/farmacología , Tiempo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/fisiología , Vasoconstrictores/farmacología , Vasodilatación/fisiología , Vasodilatadores/farmacologíaRESUMEN
AIM: This in vitro study using rat cortical slices, isolated proximal tubules and mitochondria was conducted to investigate the effect of exogenous and endogenous nitric oxide on ammoniagenesis. METHODS AND RESULTS: The cortical slices were incubated with phosphate-buffered saline containing 1 mML-glutamine at 37 degrees C andglutamine-stimulated ammoniagenesis which was further elevated with 10(-7)M ANGII showed a time-dependent decrease during 2 h. 10(-4)M L-NAME or 10(-5)ML-canavanin caused a similar ammonia elevation to that of ANGII, whereas the addition of 10(-5)M SNAP attenuated the ammonia-increasing effects of ANGII and L-NAME. Basal or exogenous NO without significantly affecting glutamine uptake of the slices seemed to convert the glutamine deamidation pathway to transamination, since L-NAME increased the ammonia to glutamine ratio from 0.87 +/- 0.08 mol/mol to 1.03 +/- 0.04 (p < 0.01). L-NAME increased both ammoniagenesis and mitochondrial oxygen consumption but SNAP depressed them. Endogenous NO reduced ammoniagenesis without changing the mitochondrial permeability transition pore (PTP), whereas exogenous NO-induced attenuation in ammoniagenesis was associated with elevated PTP in a CsA-sensitive manner. CONCLUSION: These results demonstrated that in rat kidney, basal NO depresses mitochondrial oxygen consumption and attenuates ammoniagenesis without affecting PTP; however, exogenous NO inhibits ammonia production by disturbing PTP in isolated mitochondria.
Asunto(s)
Amoníaco/metabolismo , Corteza Renal/metabolismo , Túbulos Renales Proximales/metabolismo , Mitocondrias/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico/farmacología , Oxígeno/metabolismo , Animales , Permeabilidad de la Membrana Celular/efectos de los fármacos , Permeabilidad de la Membrana Celular/fisiología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Corteza Renal/citología , Corteza Renal/efectos de los fármacos , Túbulos Renales Proximales/diagnóstico por imagen , Túbulos Renales Proximales/efectos de los fármacos , Masculino , Mitocondrias/efectos de los fármacos , Consumo de Oxígeno/efectos de los fármacos , Ratas , UltrasonografíaRESUMEN
In order to reduce the haemolytic susceptibility of glucose-6-phosphate dehydrogenase (G-6-PD) deficient erythrocytes, Ginkgo biloba extract (EGb 761) and vitamin E (vit E) were used as antioxidant agents and their effects compared. The erythrocyte suspensions from control and G-6-PD deficient patients were subjected to hydrogen peroxide (H2O2) incubation for 1 h. The produced thiobarbituric acid reactive substance (TBARS) levels measured as nmol/g Hb were compared with those of the erythrocytes administered 250 microg/mL EGb 761 or vit E previously or concomitantly with H2O2. Preincubation with EGb 761 reduced the TBARS levels from 317.14 +/- 25.27 to 160.09 +/- 21.97 nmol/g Hb in controls and from 348.24 +/- 7.79 to 205.60 +/- 14.22 nmol/g Hb in deficient erythrocytes. Concomitant application of EGb 761 with H2O2 resulted in similar but less reduction. The antioxidative effects of vitamin E were comparable to those of EGb 761. Contrary to the results obtained from oxidant conditions, the antioxidant characteristics of EGb 761 and vitamin E were not observed when they were applied directly to the erythrocytes without oxidative stress. The findings demonstrate that irrespective of administration time, EGb 761 significantly reduced TBARS levels in the erythrocytes of control and G-6-PD deficient patients subjected to oxidative stress.
Asunto(s)
Antioxidantes/farmacología , Eritrocitos/efectos de los fármacos , Ginkgo biloba , Peroxidación de Lípido/efectos de los fármacos , Fitoterapia , Extractos Vegetales/farmacología , Adolescente , Adulto , Antioxidantes/administración & dosificación , Antioxidantes/uso terapéutico , Estudios de Casos y Controles , Niño , Preescolar , Eritrocitos/metabolismo , Deficiencia de Glucosafosfato Deshidrogenasa/sangre , Humanos , Peróxido de Hidrógeno , Masculino , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
Cadmium (Cd) toxicity was produced in male rats to study the role of cholinoceptors in Cd-induced endothelial dysfunction. The changes in the tension of the aortic rings to constrictor and dilator agonists were compared with those of controls. A Cd-induced significant increase in phenylephrine response was associated with a decrease in basal dilator prostanoid release. In Cd-exposed rings, despite an obvious depression in the acetylcholine (ACh) response, the receptor-independent dilation to the calcium ionophore A23187, which elicits a receptor-independent endothelial relaxation, was slightly elevated (p<0.01), but the smooth muscle cell response to the NO donor, sodium nitroprusside (SNP) remained unaltered. Cadmium decreased both the maximal response to ACh (10(-5) M) and its pirenzepine (Prz) sensitive component. The M1 type cholinoceptor-mediated response to ACh decreased in Cd-exposed rings to 10.30 +/- 5.00% from 38.40 +/- 6.90% (p<0.001). Cadmium also reduced the share of indomethacin 1.64% to 13.92 +/- 2.89% (p<0.01), which correlated well with the changes in the M1-mediated response (r=0.991, p<0.0001). Most of the deleterious effect of Cd appears to be restricted to the M1-dependent ACh response. These findings suggest that Cd produces an endothelial dysfunction by impairing the M1 type cholinoceptor mediated response, which seems to be involved in prostanoid release.
Asunto(s)
Cadmio/toxicidad , Endotelio Vascular/fisiopatología , Enfermedades Vasculares Periféricas/inducido químicamente , Receptor Muscarínico M1/efectos de los fármacos , Receptor Muscarínico M1/fisiología , Acetilcolina/administración & dosificación , Acetilcolina/antagonistas & inhibidores , Acetilcolina/farmacocinética , Administración Oral , Animales , Aorta Torácica , Atropina/administración & dosificación , Atropina/farmacocinética , Cadmio/administración & dosificación , Cadmio/sangre , Calcimicina/administración & dosificación , Calcimicina/farmacocinética , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Endotelio Vascular/efectos de los fármacos , Trietyoduro de Galamina/administración & dosificación , Trietyoduro de Galamina/farmacocinética , Tasa de Filtración Glomerular/efectos de los fármacos , Hipertensión/inducido químicamente , Hipertensión/complicaciones , Indometacina/administración & dosificación , Indometacina/farmacocinética , Corteza Renal/química , Corteza Renal/efectos de los fármacos , Corteza Renal/fisiopatología , Enfermedades Renales/inducido químicamente , Enfermedades Renales/complicaciones , Masculino , Contracción Muscular/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , NG-Nitroarginina Metil Éster/administración & dosificación , NG-Nitroarginina Metil Éster/farmacocinética , Nitroprusiato/administración & dosificación , Nitroprusiato/farmacocinética , Enfermedades Vasculares Periféricas/complicaciones , Enfermedades Vasculares Periféricas/fisiopatología , Fenilefrina/administración & dosificación , Fenilefrina/farmacocinética , Pirenzepina/administración & dosificación , Pirenzepina/farmacocinética , Prostaglandinas/metabolismo , Ratas , Ratas Wistar , Vasodilatación/efectos de los fármacosRESUMEN
Because zinc attenuates endothelial cell dysfunction that proceeds atherosclerosis, depressed zinc status may be involved in the initiation of endothelial dysfunction. However, before recommending a zinc-enriched diet to reduce the risks for atherosclerosis, the effect of excess zinc on endothelial cell functions in normozincemic status should be known. Therefore, in this study, the effect of dietary zinc on normal endothelial cell functions in animals subjected to a diet containing 334 +/- 58 ppm zinc for 30 d was studied to see whether supplemented zinc has an effect on endothelial cells. Despite a slight increase in blood zinc, unaltered aortic and kidney zinc contents were associated with unchanged blood pressure in rats subjected to a zinc-enriched diet. Increased basal nitric oxide and prostacyclin were accompanied by a normal response to phenylephrine. Dietary zinc influenced neither endothelial-dependent nor endothelial-independent relaxations significantly. However, it elevated the share of M1-type cholinoceptor response as well as dilator prostaglandin release, which seems to be nitric oxide dependent. There was a strong correlation (r=0.826, p<0.05) between M1-type cholinoceptor response and prostacyclin release in zinc-treated rings. These results suggested that zinc ions increases M1-mediated prostacyclin release in normal endothelial cells without altering intracellular pathways.
Asunto(s)
Dieta , Endotelio Vascular/fisiología , Vasodilatación/efectos de los fármacos , Zinc/farmacología , Acetilcolina/farmacología , Administración Oral , Animales , Aorta/efectos de los fármacos , Aorta/fisiología , Calcimicina/farmacología , Relación Dosis-Respuesta a Droga , Endotelio Vascular/efectos de los fármacos , Epoprostenol/farmacología , Masculino , Nitroprusiato/farmacología , Fenilefrina/farmacología , Ratas , Ratas Wistar , Vasodilatación/fisiología , Zinc/administración & dosificaciónRESUMEN
We studied the effect of hypercholesterolemia on the pressure-sensing and regulating ability of the kidneys, using an acute hemorrhage model to provide 40% and 60% reduction in the blood pressure of hypercholesterolemic and control rats. The control group (n = 22) was fed a normal rat pellet diet and tap water; the experimental group (n = 22) received a diet containing 2% cholesterol/0.2% thaurocholate. Half the animals were subjected to 6 mL/kg bw and the others to 12 mL/kg bw of bleeding for 1 min. Blood pressure recording and proper samplings were done before bleeding and during the 20 min post-hemorrhagic period for analysis. Despite a finding of hypercholesterolemia in the experimental group, kidney cholesterol content as well as its function remained unchanged. Following an initial 40% decrease in rats bled 6 mL/kg bw, 20 min later the mean blood pressure returned to 90% of its initial value in control rats and to 70% of its basal level in hypercholesterolemic rats. A similar delay in pressure normalization occurred in rats subjected to 12 mL/kg of bleeding. Plasma renin activity remained unaffected. We conclude that dietary hypercholesterolemia delays the normalization of blood pressure after hemorrhage without affecting the sensing ability of kidneys, and that the kidneys are less sensitive than other organs to plasma cholesterol levels.
Asunto(s)
Colesterol/farmacología , Colesterol/fisiología , Riñón/efectos de los fármacos , Riñón/fisiología , Administración Oral , Animales , Presión Sanguínea/efectos de los fármacos , Colesterol/química , Colesterol en la Dieta/administración & dosificación , Modelos Animales de Enfermedad , Esquema de Medicación , Membrana Eritrocítica/química , Hipercolesterolemia/inducido químicamente , Hipotensión/complicaciones , Hipotensión/fisiopatología , Riñón/química , Hígado/química , Hígado/efectos de los fármacos , Masculino , Fosfolípidos/sangre , Fosfolípidos/química , Ratas , Renina/sangre , Renina/fisiología , Choque Hemorrágico/complicaciones , Choque Hemorrágico/fisiopatología , Ácido Taurocólico/farmacología , Factores de TiempoRESUMEN
Cadmium, which has adverse effects on many physiological systems, is an important environmental pollutant. Our previous experimental study showed that cadmium also has a dose-dependent deleterious effect on the auditory system in rats. Because zinc reverses cadmium cytotoxicity in many systems, we investigated the possible preventive effect of a zinc-enriched diet given isochronally on cadmium-induced hearing loss in rats. Fifty-four male rats were divided into three equal groups. Control rats were fed normal rat food and tap water, whereas the cadmium group was subjected to 15 ppm cadmium-containing water as CdCl2. The third group received 15 ppm CdCl2 and food enriched with 200 ppm zinc as ZnSO4 for 30 d. On d 30, eight animals from each group were used for the measurement of kidney functions. In the remaining animals, hearing functions were measured by auditory brainstem response and distortion product otoacoustic emission. Blood cadmium increased from 1.87+/-1.69 to 6.08+/-2.62 microg/dL and elevated cadmium contents of ear ossicles and kidney cortex were associated with a decreased glomerular filtration rate in rats subjected to high cadmium. A zinc-enriched diet obviously reduced cadmium accumulation in the kidney and prevented the nephrotoxicity. Our data indicated that cadmium-induced ototoxicity seems to be partially zinc preventable and zinc addition to diet without altering cadmium content in ear ossicles may help to prevent cadmium-induced hearing loss.