The enzyme kinetics of cardiac Na(+)/K(+)-ATPase were used for characterizing the ATP- and Na(+)-binding sites after administration of red wine polyphenolic compounds (Provinol) during developing and sustained hypertension. Hypertension was induced in rats (LN group) by the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 40 mg*kg(-1)*day(-1)). Provinol (40 mg*kg(-1)*day(-1)) was applied during developing hypertension (LNPF4 group) and sustained hypertension (LNPF7/3 group). Provinol reduced the number of active Na(+)/K(+)-ATPase molecules in cardiac tissue, as indicated by decreased V(max) values (by 33% in LNPF4 and 26% in LNPF7/3 compared with LN). Concerning qualitative properties of the enzyme, Provinol induced different effects on the ATP- and Na(+)-binding sites of Na(+)/K(+)-ATPase. The ATP-binding site was impaired by Provinol, as indicated by increased K(m) value (by 52% in LNPF4 vs. LN), suggesting worsened utilization of substrate by the enzyme. In sustained hypertension, however, Provinol had no effect on the ATP-binding site, as indicated by unchanged K(m) value (LNPF7/3 vs. LN). On the other hand, the Na(+)-binding site was protected by Provinol, as suggested by decreased K(Na) value (by 72% in LNPF4 and 69% in LNPF7/3 vs. LN), indicating an increased affinity of the enzyme for sodium. Thus, Provinol appeared to stimulate the extrusion of Na(+) from cardiac cells, especially in the physiologically important range of sodium concentrations (2-10 mmol*L(-1)), during both developing and sustained hypertension.