大鼠肥大细胞中的钙 - pH 相互作用：胞质碱化而非细胞内钙释放是脱颗粒的充分信号。
Calcium-pH crosstalks in rat mast cells: cytosolic alkalinization, but not intracellular calcium release, is a sufficient signal for degranulation.
作者信息
Alfonso A, Cabado A G, Vieytes M R, Botana L M
机构信息
Departamento de Farmacologìa, Facultad de Veterinaria, 27002 Lugo, Spain.
出版信息
Br J Pharmacol. 2000 Aug;130(8):1809-16. doi: 10.1038/sj.bjp.0703490.
Abstract
The aim of this work was to study the relationship between intracellular alkalinization, calcium fluxes and histamine release in rat mast cells. Intracellular alkalinization was induced by nigericin, a monovalent cation ionophore, and by NH(4)Cl (ammonium chloride). Calcium cytosolic and intracellular pH were measured by fluorescence digital imaging using Fura-2-AM and BCECF-AM. In rat mast cells, nigericin and NH(4)Cl induce a dose-dependent intracellular alkalinization, a dose-dependent increase in intracellular calcium levels by releasing calcium from intracellular pools, and an activation of capacitative calcium influx. The increase in both intracellular calcium and pH activates exocytosis (histamine release) in the absence of external calcium. Under the same conditions, thapsigargin does not activate exocytosis, the main difference being that thapsigargin does not alkalinize the cytosol. After alkalinization, histamine release is intracellular-calcium dependent. With 2.5 mM EGTA and thapsigargin the cell response decreases by 62%. The cytosolic alkalinization, in addition to the calcium increase it is enough signal to elicit the exocytotic process in rat mast cells.
摘要
这项工作的目的是研究大鼠肥大细胞内碱化、钙通量与组胺释放之间的关系。尼日利亚菌素（一种单价阳离子载体）和氯化铵可诱导细胞内碱化。使用Fura-2-AM和BCECF-AM通过荧光数字成像测量细胞溶质钙和细胞内pH值。在大鼠肥大细胞中，尼日利亚菌素和氯化铵可诱导剂量依赖性的细胞内碱化、通过从细胞内钙库释放钙导致细胞内钙水平呈剂量依赖性增加以及钙池操纵性钙内流的激活。在无细胞外钙的情况下，细胞内钙和pH值的增加均可激活胞吐作用（组胺释放）。在相同条件下，毒胡萝卜素不激活胞吐作用，主要区别在于毒胡萝卜素不会使细胞质碱化。碱化后，组胺释放依赖于细胞内钙。加入2.5 mM乙二醇双四乙酸（EGTA）和毒胡萝卜素后，细胞反应降低62%。除了钙增加外，细胞质碱化足以作为引发大鼠肥大细胞胞吐过程的信号。
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