Abstract

Phenol red (PR) is used as a pH indicator in cell culture medium. We found that cell culture medium containing PR relaxed canine lingual arteries (LA) contracted by the thromboxane A2/prostaglandin endoperoxide (TPE) receptor agonist (15S)-hydroxy-11-alpha-9-alpha- (epoxymethano)prosta-5Z,13E-dienoic acid (U46619). We tested the effect of PR and the TPE receptor antagonist ONO-3708 on U46619, prostaglandin F2 alpha (PGF2 alpha), phenylephrine (PE) and potassium chloride (KCl)-induced contraction of the LA and on human platelet aggregation to U46619, ADP, arachidonic acid (AA), A23187 and thrombin. U46619, PGF2 alpha, KCl and PE produced equal tension development of the LA. PR relaxed the LA contracted with U46619 and PGF2 alpha with IC50 concentrations of 18.3 +/- 10 and 37.3 +/- 8.8 microM, respectively. ONO-3708 inhibited the contractions to U46619 and PGF2 alpha with IC50 of 9.4 +/- 2.2 and 12.2 +/- 2.2 nM, respectively. However, PR (300 microM) and ONO-3708 (300 nM) did not affect contraction of the LA to KCl or PE. PR inhibited human platelet aggregation, in vitro, to AA and U46619 and second wave aggregation to ADP but did not affect thrombin or first wave ADP-mediated platelet aggregation. PR inhibited U46619 and AA-induced changes in cyclic AMP and Fura-2 calcium transients in platelets and LA. However, PR did not affect the activation of cyclic AMP or intracellular calcium ion in platelets or calcium influx and the release of intracellular calcium ion in canine LA produced by ryanodine, KCl and PE. The concentration of PR in many culture media is between 40 and 70 microM. The data support the conclusion that PR, in concentrations used as a pH indicator, is a selective antagonist of TPE receptors.