Ishihara H, Shimura S, Sato M, Masuda T, Ishide N, Miura M, Sasaki T, Sasaki H, Takishima T
First Department of Internal Medicine, Tohoku University School of Medicine, Sendai, Japan.
Am J Physiol. 1990 Dec;259(6 Pt 1):L345-50. doi: 10.1152/ajplung.1990.259.6.L345.
We measured the intracellular free calcium ion concentration [( Ca2+]i) of acinar cells in isolated feline tracheal submucosal glands in response to secretagogues using the Ca2(+)-sensitive fluorescent dye fura-2. The secretagogues included cholinergic, adrenergic agonists, substance P (SP), and vasoactive intestinal polypeptide (VIP) which induce mucus glycoprotein secretion from feline tracheal submucosal glands. Methacholine (MCh) produced a significant increase in [Ca2+]i of up to 9.8 times that of control in a dose-dependent fashion at concentrations of 10(-8) to 10(-3) M. [Ca2+]i increase by MCh reached a peak within 30 s after stimulation and thereafter showed a sustained rise. In Ca2(+)-free medium, MCh produced an initial transient rise, which was less than 30% of that in a Ca2(+)-containing solution, and which lasted for 60 s with no prolonged sustained rise in [Ca2+]i. Atropine abolished MCh-evoked [Ca2+]i increase. Phenylephrine and SP produced a prolonged increase in [Ca2+]i without an initial transient increase. Phenylephrine (up to 10(-4) M) evoked an increase in [Ca2+]i by up to 240% that of control, which was abolished by prazosin. SP (up to 10(-4) M) also evoked an increase in [Ca2+]i by 155% that of control, which was abolished by atropine. By contrast, both isoproterenol (up to 10(-5) M) and VIP (up to 10(-5) M) failed to alter [Ca2+]i. These findings indicate that the mucus glycoprotein secretion evoked by muscarinic cholinergic, alpha-adrenergic agonist or SP can be mediated by intracellular Ca2+, whereas that by beta-adrenergic agonists or VIP cannot.
我们使用钙离子敏感荧光染料fura-2,测量了分离的猫气管黏膜下腺泡细胞对促分泌剂产生反应时的细胞内游离钙离子浓度[Ca2+]i。促分泌剂包括胆碱能、肾上腺素能激动剂、P物质(SP)和血管活性肠肽(VIP),它们可诱导猫气管黏膜下腺分泌黏液糖蛋白。在浓度为10(-8)至10(-3) M时,乙酰甲胆碱(MCh)以剂量依赖方式使[Ca2+]i显著增加,最高可达对照的9.8倍。MCh刺激后,[Ca2+]i在30秒内达到峰值,随后持续升高。在无钙培养基中,MCh产生初始短暂升高,幅度小于含钙溶液中的30%,持续60秒,[Ca2+]i无延长的持续升高。阿托品消除了MCh引起的[Ca2+]i升高。去氧肾上腺素和SP使[Ca2+]i持续升高,无初始短暂升高。去氧肾上腺素(最高10(-4) M)使[Ca2+]i升高至对照的240%,哌唑嗪可消除该作用。SP(最高10(-4) M)也使[Ca2+]i升高至对照的155%,阿托品可消除该作用。相比之下,异丙肾上腺素(最高10(-5) M)和VIP(最高10(-5) M)均未改变[Ca2+]i。这些发现表明,毒蕈碱型胆碱能、α-肾上腺素能激动剂或SP引起的黏液糖蛋白分泌可由细胞内钙离子介导,而β-肾上腺素能激动剂或VIP引起的则不能。