Irvine F, Pyne N J, Houslay M D
FEBS Lett. 1986 Nov 24;208(2):455-9. doi: 10.1016/0014-5793(86)81068-7.
Treatment of intact hepatocytes with the phorbol ester 12-O-tetradecanoyl phorbol 13-acetate (TPA) potentiated the ability of glucagon to increase intracellular cyclic AMP concentrations. This effect was dose-dependent upon TPA, exhibiting an EC50 of 0.39 ng/ml and such activation was observed at both saturating and sub-saturating concentrations of glucagon. However, this stimulatory effect of TPA was completely abolished by the presence of the cyclic AMP phosphodiesterase inhibitor 1-isobutyl-3-methylxanthine, when TPA now inhibited the glucagon-stimulated increase in intracellular cyclic AMP concentrations. It is suggested that, as well as inhibiting glucagon-stimulated adenylate cyclase activity, TPA also inhibits cyclic AMP phosphodiesterase activity in intact hepatocytes. Treatment of either hepatocyte homogenates or purified cyclic AMP phosphodiesterase with TPA failed to show any direct inhibitory effect of TPA on activity showing that TPA did not exert any direct inhibitory action on phosphodiesterase activity. However, homogenates made from hepatocytes that had been pre-treated with TPA did show a reduced cyclic AMP phosphodiesterase activity. It is suggested that TPA might inhibit cyclic AMP phosphodiesterase activity through phosphorylation by C-kinase.
用佛波酯12 - O -十四烷酰佛波醇13 - 乙酸酯(TPA)处理完整的肝细胞,可增强胰高血糖素提高细胞内环磷酸腺苷(cAMP)浓度的能力。这种效应呈TPA剂量依赖性,半数有效浓度(EC50)为0.39 ng/ml,并且在胰高血糖素的饱和及亚饱和浓度下均观察到这种激活作用。然而,当存在环磷酸腺苷磷酸二酯酶抑制剂1 -异丁基- 3 -甲基黄嘌呤时,TPA的这种刺激作用完全被消除,此时TPA反而抑制了胰高血糖素刺激引起的细胞内环磷酸腺苷浓度升高。提示TPA除了抑制胰高血糖素刺激的腺苷酸环化酶活性外,还抑制完整肝细胞中的环磷酸腺苷磷酸二酯酶活性。用TPA处理肝细胞匀浆或纯化的环磷酸腺苷磷酸二酯酶,均未显示TPA对其活性有任何直接抑制作用,表明TPA对磷酸二酯酶活性未产生任何直接抑制作用。然而,用TPA预处理过的肝细胞制成的匀浆确实显示出环磷酸腺苷磷酸二酯酶活性降低。提示TPA可能通过蛋白激酶C磷酸化来抑制环磷酸腺苷磷酸二酯酶活性。