On the mechanisms of the antispasmodic action of some hindered phenols in rat aorta rings

Abstract

The antispasmodic effects of 3- t-butyl-4-hydroxyanisole (BHA) and some structurally related compounds were investigated in endothelium-intact rat aorta rings. Nordihydroguaieretic acid (NDGA), BHA, 3,5-di- t-butyl-4-hydroxyanisole (DTBHA), 2,6-di-isopropyl phenol (propofol) and 2,2′-dihydroxy-3,3′-di- t-butyl-5,5′-dimethoxydiphenyl (DIBHA) did not cause relaxation when added at the plateau of phenylephrine-evoked contraction, nor did they affect the concentration–relaxation curve for acetylcholine in precontracted rings. In rings depolarised with physiological salt solution (PSS) containing 40 mM K +, NDGA, BHA, DTBHA, 2,5-di- t-butyl-1,4-benzohydroquinone (BHQ), propofol and nifedipine, but not DIBHA, inhibited the contraction induced by cumulative addition of Ca 2+ (0.05–10 mM) in a concentration-dependent manner; this inhibition was inversely related to the Ca 2+ concentration. In 40 mM K + PSS, 25 nM nifedipine blocked the 1 mM Ca 2+-induced contraction, whereas 50 μM DTBHA, NDGA, BHA, BHQ and propofol significantly antagonised it by 84.4%, 73.0%, 52.8%, 45.6% and 35.7%, respectively. In the presence of 1 μM methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2-trifluoromethylphenyl)-pyridine-5-carboxylate (Bay K 8644), the response to Ca 2+ did not differ from control values with nifedipine and BHQ, was partially restored with DTBHA and NDGA, and was not affected with BHA and propofol. Nifedipine markedly inhibited (85.2%) the Ba 2+-induced contraction and this effect was totally reversed by Bay K 8644. BHA and DTBHA showed antispasmodic activity (45.3% and 43.1%, respectively) which was partly reversed by Bay K 8644. In contrast, Bay K 8644 did not affect the inhibition exerted by BHQ, NDGA and propofol (69.5%, 53.3% and 46.1%, respectively). Nifedipine, BHA, DTBHA, propofol and NDGA inhibited the contractile response to 1 mM Ca 2+ of aorta rings depolarised with 40 or 80 mM K + PSS to a similar extent. Cromakalim inhibited the Ca 2+-evoked contraction only in 30 mM K + PSS and BHQ only in 80 mM K + PSS. DIBHA had no effect on this model. Cromakalim, but not BHA, stimulated 86Rb + efflux from ring preparations. In 80 mM K + PSS containing 1 μM nifedipine, only papaverine affected the phenylephrine-induced contraction. Moreover, when the rings were preincubated with 1 mM Ni 2+, the response to phenylephrine in the presence of BHQ was significantly reduced. In conclusion, we propose that BHA may non-specifically inhibit Ca 2+ influx at the plasmalemma level rather than affect the function of K + channels, Ca 2+ release from intracellular stores or endothelium-dependent relaxation.