Angiotensin type-1 (AT1) receptor gene expression in primarily cultured human arterial umbilical endothelial cells

The aim of this study was to investigate the interaction of angiotensin II (Ang II) and human umbilical arterial endothelial cells (HUAEC). Specific binding of 125I-ANG II to primarily cultured HUAEC showed a K(D) of 1.98 +/- 0.53 x 10(-9) M (n = 5) with a maximum binding site of 2.84 +/- 1.07 x 10(-13) mol/mg protein (n = 5). In later passages (third and fifth subculture), this binding site was no longer detectable. Gene expression analysis revealed a strong expression of the angiotensin type-I receptor (AT1-R) in the primarily cultured HUAEC, with a decrease in additional passages. In primarily cultured HUAEC, Ang II (10(-10)-10(-6) M) induced a concentration-dependent increase in intracellular free calcium ([Ca2+]i) that could be blocked by a preincubation with candesartan (TCV-112) but not by PD123319. These data show the expression of the AT1-R in primary cultures of HUAEC. The Ang II-induced increase in [Ca2+]i seems to be mediated by this receptor.