Insulin stimulates the Na+,K(+)-ATPase and the Na+/K+/Cl- cotransporter of human fibroblasts.
Insulin regulation of K+ (Rb+) transport was investigated in cultured human fibroblasts using a non-radioactive method which allows the simultaneous determination of the intracellular concentration of other monovalent cations. Insulin stimulated Rb+ influx through the Na+,K(+)-ATPase and the Na+/K(+)/Cl- cotransporter in human fibroblasts. Insulin stimulation was very rapid and maximal effect was observed within 10 min. Insulin stimulation of Rb+ uptake via the Na+,K(+)-ATPase and the Na+/K(+)/Cl- cotransporter was dose-dependent, with half-maximal stimulation at 2-3 nM of hormone. Insulin increased the V(max) of both transporters involved, affecting only minimally their Km. In other cells, insulin stimulates the Na+,K(+)-pump by increasing Na+ availability through the Na+/H+ exchanger. In human fibroblasts, insulin stimulation of Na+,K(+)-ATPase occurred in the presence of ethyl-isopropyl amiloride, an inhibitor of the Na+/H+ exchanger, and without sustained changes in intracellular[Na+]. By contrast, insulin action on Na+,K(+)-ATPase was impaired by the protein kinase inhibitors staurosporine and genistein. These results indicate that, in human fibroblasts, insulin stimulates both the Na+,K(+)-ATPase and the Na+/K+/Cl- cotransporter, that stimulation of the Na+,K(+)-ATPase occurs in the absence of changes in intracellular [Na+], and that protein kinase activity is essential for this insulin action.
Longo
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Department of Pediatrics, Emory University, Atlanta, GA 30322, USA.
D000328:Adult; D002034:Bumetanide / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D005347:Fibroblasts; D006801:Humans; D007328:Insulin / Q000494:pharmacology*; D007700:Kinetics; D010042:Ouabain / Q000494:pharmacology; D047428:Protein Kinase Inhibitors; D012413:Rubidium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D028021:Sodium-Potassium-Chloride Symporters; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
D002352:Carrier Proteins; D007328:Insulin; D047428:Protein Kinase Inhibitors; D028021:Sodium-Potassium-Chloride Symporters; D002034:Bumetanide; D010042:Ouabain; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase; D012413:Rubidium