Thrombin causes endothelium-dependent biphasic regulation of vascular tonein the porcine renal interlobar artery

1 Using a method employing front-surface fura-2 fluorometry to measure the cytosolic Ca2+ concentration, [Ca2+](i), the mechanism of endothelium-depen dent regulation of vascular tone by thrombin was studied in porcine renal i nterlobar arterial strips. 2 At concentrations lower than 3 u ml(-1), thrombin evoked only early trans ient relaxation, while at 3 u ml(-1) and higher concentrations, thrombin ca used an early relaxation and a subsequent transient contraction. Both throm bin-induced relaxation and contraction were abolished by removing the endot helium. Similar biphasic responses were observed with a protease-activated receptor-1-activating peptide. 3 Early relaxation was associated with a decrease in [Ca2+](i), while the t ransient contraction was not associated with a change in [Ca2+](i) of smoot h muscle cells. 4 A thromboxane A(2) (TXA(2))/prostaglandin H-2 (PGH(2)) receptor antagonis t (10(-5) M ONO-3708) completely inhibited the thrombin-induced contraction , whereas a thromboxane Aa synthase inhibitor (10(-5) M OKY-046) only partl y inhibited it. 5 When the thrombin-induced contraction was inhibited by ONO-3708, either p retreatment with N-omega-nitro-L-arginine methylester (L-NAME) or an increa se in the amount of external K+ to 40 mM did not abolish thrombin-induced r elaxation during phenylephrine-induced sustained contraction. However, the combination of pretreatment with L-NAME and an elevation of external K+ to 40 mM completely abolished the relaxation. 6 There was no significant difference in the concentration-dependent effect s of thrombin on the initial early relaxation between conditions in which t he contractile components either were or were not inhibited. 7 Thrombin is thus considered to mainly activate protease-activated recepto r-1 and cause a biphasic response, early relaxation and a transient contrac tion, in the porcine renal interlobar artery in an endothelium-dependent ma nner. The thrombin-induced endothelium-dependent relaxation was mediated by nitric oxide and hyperpolarizing factors, while the contraction was mediat ed by TXA(2) and PGH(2).