HISTAMINE-INDUCED CALCIUM RELEASE AND PHORBOL ANTAGONISM IN CULTURED AIRWAY SMOOTH-MUSCLE CELLS
HISTAMINE-INDUCED CALCIUM RELEASE AND PHORBOL ANTAGONISM IN CULTURED AIRWAY SMOOTH-MUSCLE CELLS
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DOI：
10.1152/ajpcell.1987.253.4.c561
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发表时间：
1987-10-01
影响因子：
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通讯作者：
REYNOLDS, EE
中科院分区：
文献类型：
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作者：
KOTLIKOFF, MI;MURRAY, RK;REYNOLDS, EE
将原代培养的气道平滑肌细胞暴露于组胺，并用钙敏感活性染料Fura-2测定细胞内游离钙瞬变。100微米组胺刺激导致细胞内钙离子从未刺激的178+-水平升高。25至497+-154 nM钙离子(SE；n=14)，并在刺激后1分钟内恢复到基线游离钙浓度。用H1受体阻滞剂吡拉明(2.5微米)预处理细胞可消除这种反应；然而，用H2受体阻滞剂西咪替丁(50微米)预处理，或用2 mM的CO~(2+)或5微米硝苯地平预处理，不能改变细胞的钙瞬变。200 nM佛波醇12-肉豆蔻酸盐13-乙酸酯(PMA)激活蛋白激酶c后，胞浆内钙无明显升高，但完全阻断组胺对细胞内钙的释放。我们得出的结论是：1)组胺引起气道平滑肌细胞内钙的一过性升高，2)细胞内钙的升高是由H1受体偶联引起的，3)蛋白激酶c的激活阻断了组胺诱导的细胞内钙的释放。
Primary cultures of airway smooth muscle cells were exposed to histamine, and intracellular free calcium transients were measured by the calcium-senstivitve dye fura-2. Stimulation with 100 .mu.M histamine resulted in a rise in intracellular calcium from an unstimulated level of 178 .+-. 25 to 497 .+-. 154 nM Ca2+ (SE; n = 14) and a return to base-line free calcium concentration within 1 min of stimulation. Pretreatment of cells with the H1 receptor blocker pyrilamine (2.5 .mu.M) abolished the response; however, the calcium transient was not altered by pretreatment with the H2 blocker cimetidine (50 .mu.M), by chelation of external calcium, or by pretreatment with 2 mM Co2+ or 5 .mu.M nifedipine. Activation of protein kinase c by 200 nM phorbol 12-myristate 13-acetate (PMA) resulted in no detectable rise in cytosolic calcium but completely blocked the release of internal calcium by histamine. We conclude that 1) histamine causes a transient rise of cytosolic calcium in airway smooth muscle, 2) the rise in cytosolic calcium is mediated by H1 receptor coupling that triggers release of internal calcium stores, and 3) activation of protein kinase c blocks the histamine-induced release of intracellular calcium.