Isosteviol as a potassium channel opener to lower intracellular calcium concentrations in cultured aortic smooth muscle cells

Isosteviol is a derivative of stevioside, a constituent of Stevia rebaudiana, and is commonly used as a non-caloric sugar substitute in Japan and Brazil. The present study attempted to elucidate the role of potassium (K ⁺) channels in the action of isosteviol on intracellular calcium concentrations ([Ca²⁺]i) in cultured vascular smooth muscle (A7r5) cells using the Ca²⁺-sensitive dye Fura-2 as an indicator. The increase of [Ca²⁺]i in A7r5 cells produced by vasopressin (1 μmol/L) or phenylephrine (1 μmol/L) was attenuated by isosteviol from 0. 01 μmol/L to 10 μmol/L. The attenuation by isosteviol of the vasopressin- and phenylephrine-induced increase in [Ca²⁺]i was inhibited by glibenclamide, apamin and 4-aminopyridine but not by charybdotoxin. Furthermore, the inhibitory action of isosteviol on [Ca²⁺]i was blocked when A7r5 cells co-treated with glibenclamide and apamin in conjunction with 4-aminopyridine were present. Therefore, not only did the ATP-sensitive potassium (K_ATP) channel affect the action of isosteviol on [Ca ²⁺]i modulation in A7r5 cells, but also those on the small conductance calcium-activated potassium (SK_Ca) channels and voltage-gated (Kv) channels. However, the blockers of large-conductance Ca ²⁺-activated potassium channels failed to modify the inhibitory action of isosteviol on [Ca²⁺]i. The obtained results indicated that a decrease of [Ca²⁺]i in A7r5 cells by isosteviol is mainly mediated by the selective opening of K_ATP channel or/and SK_Ca channel. Alteration in the Kv channel also plays a critical role in the inhibitory action of isosteviol.