Intracellular sodium, potassium and magnesium concentration, ouabain-sensitive 86rubidium-uptake and sodium-efflux and Na+, K+-cotransport activity in erythrocytes of normal male subjects studied on two occasions.

The red cell Na+,K+-ATPase pump activity estimated by the ouabain-sensitive 86Rb-uptake or Na+-efflux, the Na+,K+-cotransport activity measured either by the furosemide-sensitive K+- or Na+- efflux or by the ethacrynic acid-sensitive 86Rb-uptake as well as the intraerythrocyte concentration of sodium, potassium and magnesium were studied in 29 normal male subjects with one to three weeks interval between the first and second blood sampling. Both the red cell sodium and potassium concentration, the erythrocyte ouabain-sensitive 86Rb-uptake and Na+-efflux, the furosemide-sensitive Na+- and K+-efflux and the ethacrynic acid-sensitive 86Rb-uptake are stable over time in the same individual. The furosemide-sensitive Na+-and K+-efflux is significantly related to the ethacrynic acid-sensitive 86Rb-uptake. The intraerythrocyte Na+ concentration was negatively related to the ouabain-sensitive 86Rb-uptake, but not to the ouabain-sensitive Na+-efflux; it was, however, negatively related to the rate constant for the ouabain-sensitive Na+-efflux.

[Physiologic variability of erythrocyte concentrations and transport of sodium and potassium]. / Variabilité physiologique des concentrations et des transports érythrocytaires du sodium et du potassium.

Sodium concentrations in erythrocytes are lower in women during the luteal phase of the menstrual cycle than in women studied during the follicular phase and in men. Sodium cotransport activity is lower in women during the follicular phase than in men. Women taking oral contraceptives show no monthly variations in erythrocyte sodium concentrations. Subjects with a family history of essential hypertension have higher sodium concentrations in erythrocytes than those with a different family background, partly because of reduction in sodium cotransport activity. Negroes have marked erythrocyte sodium accumulation due to lower activity of the sodium-potassium pump and cotransport system; they also have reduced sodium-lithium counter-transport. There is no difference between negroes with or without haemoglobin S. There are no changes in erythrocyte potassium concentrations in relation to sex, menstrual phase, race, familial essential hypertension, presence of haemoglobin S or use of oral contraceptives. These physiological variations indicate the factors which must be standardized to study sodium concentrations in cells and sodium transmembrane flux.

Intra-erythrocyte concentration and transmembrane fluxes of sodium and potassium during acute and short-term administration of ketanserin in normal male subjects.

The effect of acute and short-term administration of ketanserin on the intracellular concentration and transmembrane fluxes of sodium and potassium in erythrocytes was studied in 12 sodium-replete, normal male subjects. The subjects received 40 mg ketanserin three times a day for one week. Blood samples were drawn before and 2.5 hours after the first dose, 12 hours after the evening dose of the sixth day and 1.5 hours after the morning dose of the seventh day. The intra-erythrocyte sodium concentration was not changed following the first dose of ketanserin, but was decreased (p less than 0.05) during short-term treatment with ketanserin. The ouabain-sensitive 86Rb-uptake, an estimate of the Na+,K+-ATPase pump activity, was decreased (p less than 0.001) after acute ketanserin administration, but not (p = 0.07) during short-term treatment. This change in intra-erythrocyte sodium concentration was related (r = 0.73, p less than 0.01) to the change in ouabain-sensitive 86Rb-uptake. The red cell Na+,K+-cotransport and Na+,Li+-countertransport activity were not changed during acute and short-term administration of ketanserin. The results indicate that short-term ketanserin administration decreases the intra-erythrocyte sodium concentration, but the flux measurements cannot explain this observation.

Assessment of sodium-potassium ATPase activity in human erythrocytes in vitro.

The red cell Na+,K+-ATPase pump activity was measured either by the ouabain-sensitive 86Rb-uptake in the presence or absence of 4 mM potassium in the external medium or by the ouabain-sensitive Na+-efflux in normal male subjects without any medication (n = 14) and during short-term treatment with the serotonin-antagonist, ketanserin. The ouabain-sensitive 86Rb-uptake in a medium containing 4 mM K+ was related to the 86Rb-uptake in a K+-free medium (r = 0.82; p less than 0.001) and to the Na+ efflux (r = 0.74; p less than 0.01). The intra-erythrocyte sodium concentration was negatively related to the ouabain-sensitive 86Rb-uptake (K+-free medium r = 0.59, p less than 0.05: 4 mM K+ in medium r = -0.85; p less than 0.01) and to the ouabain-sensitive Na+ efflux (r = -0.53; p less than 0.05). During serotonin antagonism with ketanserin no changes were found in the ouabain-sensitive 86Rb-uptake or Na+-efflux.

Methodological assessment of assays for intracellular concentration and transmembrane fluxes of sodium and potassium in erythrocytes of man.

The reliability of the assay of intracellular Na+ and K+ concentration; Na+,K+-ATPase activity; Na+-K+ cotransport and Na+-Li+ countertransport in erythrocytes was investigated. These assays were also applied in normal male subjects with and without family history of hypertension. The various assays are reproducible as indicated by the intra-assay variation and are stable over time as shown by the inter-assay variation. A delay between blood sampling and analysis, however, result in an increase in intracellular Na+ concentration and decreases in Na+,K+-ATPase activity; Na+-K+ cotransport and Na+-Li+ countertransport. Compared to the white normal males without family history of hypertension (n = 43) red blood cells of white normal males with such a history (n = 17) have a higher (p less than 0.01) intracellular Na+ concentration. This can be at least partly due to their lower (p less than 0.001) furosemide-sensitive Na+ efflux rate. Also, their ouabain-resistant 86Rb-uptake was lower (p less than 0.05). The potassium concentration in the red blood cells was similar in both groups.