Watson E L, Jacobson K L, DiJulio D H, Dowd F J
Department of Oral Biology, University of Washington, Seattle 98195.
Am J Physiol. 1993 Oct;265(4 Pt 1):C1061-8. doi: 10.1152/ajpcell.1993.265.4.C1061.
Carbachol (0.1-10 microM) augmented the isoproterenol-stimulated adenosine 3',5'-cyclic monophosphate (cAMP) accumulation by approximately 50% in mouse parotid acini; at carbachol concentrations > 10 microM the stimulatory trend was reduced. These effects were time dependent. In the presence of 3-isobutyl-1-methylxanthine (IBMX), the overall response to carbachol was an inhibition of the isoproterenol response. Pretreatment of acini with pertussis toxin failed to reverse this inhibition, suggesting that the effects of carbachol were not related to effects on the GTP binding protein, Gi. A-23187 mimicked the effects of carbachol on isoproterenol-stimulated cAMP accumulation in the presence and absence of IBMX. In the presence of IBMX, carbachol failed to inhibit isoproterenol-stimulated cAMP accumulation when calcium was absent from the extracellular media and depleted from intracellular stores by thapsigargin. By contrast, in the absence of IBMX, removal of calcium abolished augmentation of isoproterenol responses by low concentrations of carbachol, whereas at higher carbachol concentrations isoproterenol responses were significantly inhibited; the time to maximal cAMP accumulation was decreased approximately eightfold. The results show that the mechanisms underlying the effects of carbachol on cAMP metabolism involve both the enzymes that synthesize and degrade cAMP.
卡巴胆碱(0.1 - 10微摩尔)可使异丙肾上腺素刺激的小鼠腮腺腺泡中3',5'-环磷酸腺苷(cAMP)积累增加约50%;当卡巴胆碱浓度>10微摩尔时,刺激趋势减弱。这些效应具有时间依赖性。在存在3 - 异丁基 - 1 - 甲基黄嘌呤(IBMX)的情况下,对卡巴胆碱的总体反应是对异丙肾上腺素反应的抑制。用百日咳毒素预处理腺泡未能逆转这种抑制作用,这表明卡巴胆碱的作用与对GTP结合蛋白Gi的作用无关。在存在和不存在IBMX的情况下,A - 23187模拟了卡巴胆碱对异丙肾上腺素刺激的cAMP积累的影响。在存在IBMX的情况下,当细胞外培养基中不存在钙且用毒胡萝卜素使细胞内钙库耗尽时,卡巴胆碱未能抑制异丙肾上腺素刺激的cAMP积累。相比之下,在不存在IBMX的情况下,去除钙消除了低浓度卡巴胆碱对异丙肾上腺素反应的增强作用,而在较高卡巴胆碱浓度下,异丙肾上腺素反应受到显著抑制;达到最大cAMP积累的时间减少了约八倍。结果表明,卡巴胆碱对cAMP代谢影响的潜在机制涉及合成和降解cAMP的酶。