Lack of hypophosphatemic effect on glucose uptake, lactate and pyruvate release by the normal rat perfused hind-limb

In previous experiments, it was observed that in the perfused hind-limb of phosphorus deficient rats there was a faster utilization of glucose when compared with preparations obtained from ad-libitum fed rats but similar to that obtained from pair-weighed rats. A greater release of lactate under the influence of insulin and a marked spontaneous and insulin induced liberation of pyruvate was registered in the perfused hind-limb of the phosphorus deficient rat. Since a hypophosphatemic medium was employed, we though it would be interesting to study the effect of low perfusate Pi in normal rat hind-limb preparations and, additionally, to observe it fast changes in the level of medium orthophosphate from normal to low values and viceversa were able to determine changes in glucose uptake and lactate and pyruvate release. In one group of experiments, the hind-limb of normal rats was perfused successively with orthophosphate deprived medium for thirty minutes in each stage. In another group of experiments, the first stage was carried out with normal. Pi medium, the second with orthophosphate deprived medium, and the third with normal Pi medium; each stage lasted thirty minutes. In all the experiments there was a gradual release of orthophosphate. The glucose uptake was not modified by the fast changes in the medium Pi nor were significant differences in lactate and pyruvate release observed, which showed clear increases in all stages. It is concluded that hypophosphatemia alone is not sufficient to obtain greater increases of lactate and pyruvate release; it is also required that the hind-limb preparations be obtained from ..

Inhibition of the glucagon induced incorporation of orthophosphate into the liver by gluconeogenic blockers

Hígados aislados de ratas alimentadas, fueron perfundidos con un medio preparado a partir de solución KRB a la cual se añadieron: seroalbúmina bovina, 3g/dl; glucosa, 90 mg/dl; ortofosfato para obtener una concentración en el plasma de 1 mg/dl; 18 por ciento de glóbulos rojos de rata lavados; pH, 7.40, fase gaseosa O2 95%-CO2 5%. Flujo 36 ml/min. Temperatura, 37-C. Volumen del perfusado 70 ml. 0,5 micronCi de 32P-ortofosfato por ml. fueron añadidos al perfusado diez minutos antes de montar el hígado: el tiempo, 15 minutos después de montar el órgano, se tomó como punto cero. En los experimentos en los cuales se emplearon los bloqueadores de la neoglucogénesis ( quinolinato 1,68 mM o aminoóxi-acetato 0.2 mM) éstos fueron añadidos al perfusado al tiempo 15 min. el glucagon, cuando fue administrado, se empleó en inyección continua entre 45 y 90 min. (20 microngm). Se tomaron muestras cada 15 minutos hasta 120 minutos, que se recogieron en tubos enfriados en hielo. Glucosa en el perfusado, Pi en el plasma, 32Pi en perfusado y actividad específica del Pi plasmático fueron los parámetros determinados. Se calcularon Deltas a partir de 45 min. Los valores de glucosa y de Pi plasmático se corrigieron a partir de blancos obtenidos, haciendo circular el perfusado en ausencia del hígado y los resultados se expresaron por 10g de hígado fresco