ABSTRACT
We have recently shown that insulin attenuates angiotensin II-induced intracellular Ca(2+) mobilization in human skin fibroblasts from normotensive subjects. This study was designed to investigate the effects of angiotensin II and the interactions between insulin and angiotensin II on intracellular Ca(2+) mobilization in skin fibroblasts from patients with essential hypertension. Fibroblasts were obtained from 9 normotensives and 18 hypertensives. Spectrofluorophotometric free Ca(2+) measurement was performed in monolayers of 24-hour serum-deprived cells. Resting intracellular Ca(2+) level and angiotensin II-stimulated intracellular Ca(2+) peak were higher in fibroblasts from hypertensives compared with those from normotensives. The effect of acute insulin exposure was evaluated in fibroblasts from hypertensives subdivided on the basis of insulin sensitivity. In insulin-sensitive hypertensives, insulin significantly blunted the effects of angiotensin II on intracellular Ca(2+) response, whereas in insulin-resistant patients, insulin did not modify intracellular Ca(2+) response to angiotensin II. Pertussis toxin, a G(ialpha)-inhibitor, reduced angiotensin II-stimulated Ca(2+) peak in insulin-sensitive but not in insulin-resistant hypertensives. In conclusion, the effects of angiotensin II on intracellular Ca(2+) mobilization are more pronounced in fibroblasts from hypertensives compared with those from normotensives, and the inhibitory effect of insulin is blunted in insulin-resistant hypertensives by a G(ialpha) pertussis toxin-sensitive abnormality.
Subject(s)
Angiotensin II/pharmacology , Calcium/metabolism , Fibroblasts/metabolism , Hypertension/metabolism , Insulin Resistance , Insulin/pharmacology , Adult , Cells, Cultured , Culture Media , Cytosol/metabolism , Female , Fibroblasts/drug effects , Humans , Male , Middle Aged , Pertussis Toxin , Skin/cytology , Thapsigargin/pharmacology , Virulence Factors, Bordetella/pharmacologyABSTRACT
OBJECTIVE: To ascertain the relationships between the kinetic properties of erythrocyte Na+-H+ exchange [maximum kinetic energy (Vmax), Michaelis constant (Km) for internal H+ and Hill's coefficient], Na+-Li+ (Vmax and Km for external Na+), and metabolic parameters in normotensive controls and hypertensive subjects. MATERIALS AND METHODS: Na+-H+ exchange was measured as the Na+ influx driven by intracellular H+, and Na+-Li+ exchange as the Li+ efflux driven by extracellular Na+, in erythrocytes from normotensive (n = 59) and hypertensive (n = 93) subjects. RESULTS: In comparison with normotensives, the hypertensives had a higher Vmax for Na+-Li+ and Na+-H+ exchange, a higher Km for external Na+ for Na+-Li+ exchange and a lower reduced Hill's number for Na+-H+ exchange. Vmax values for Na+-Li+ and Na+-H+ exchange were significantly correlated, as were Km values for internal H+ for Na+-H+ exchange and Km for external Na+ for Na+-Li+ exchange. Insulin resistance and beta-cell function indices were higher in the hypertensives than the normotensives. Upon stepwise multiple regression analysis, Vmax for Na+-Li+ exchange was correlated significantly and independently with Km for external Na+ and with the insulin resistance index, while Km for external Na+ was correlated with Km for internal H+, Vmax for Na+-H+ exchange and mean blood pressure. Vmax and Hill's coefficient for Na+-H+ exchange were correlated only with mean blood pressure. CONCLUSIONS: The demonstration of functional correlations between the kinetic properties of Na+-H+ and Na+-Li+ exchange provides further evidence that erythrocyte Na+-Li+ exchange is a functioning mode of Na+-H+ exchange, which is affected by insulin resistance.
