Berlin G, Enerbäck L
Agents Actions. 1984 Apr;14(3-4):401-4. doi: 10.1007/BF01973837.
The fluorescent cationic dye berberine in combination with histamine release studies have been used to explore the different steps of the mast cell secretory process. We have previously shown that quantitation of heparin release by the binding of berberine to fixed mast cells can be used as a direct measure of release of granules. This report summarizes recent work using berberine as a vital stain demonstrating the secretory activity of mast cells. After membrane stabilization with polyethylene glycol (PEG) normal mast cells exclude the dye while mast cells stimulated to secretion with polymyxin B show a strongly fluorescent dye binding to individual cytoplasmic granules. The mean fluorescence intensity of the cell populations after vital berberine staining was compared both to heparin and histamine release. The results strongly suggest that berberine, under the vital staining conditions used, is a marker of intracellular granules that have released histamine. The vital staining method was also used to study membrane events following a polymyxin B-induced secretion. The mean fluorescence intensity decreased by 75% during the first hour after the termination of a polymyxin B stimulation while the mast cell content of histamine and heparin remained constant. The findings support the idea that the membranes are rapidly restored after mast cell secretion, permitting a selective amine release without accompanying release of heparin or other matrix components of the granules.
荧光阳离子染料黄连素结合组胺释放研究已被用于探索肥大细胞分泌过程的不同步骤。我们之前已经表明，通过黄连素与固定肥大细胞的结合来定量肝素释放，可以用作颗粒释放的直接测量方法。本报告总结了最近使用黄连素作为活体染色剂来证明肥大细胞分泌活性的工作。在用聚乙二醇（PEG）稳定细胞膜后，正常肥大细胞会排斥该染料，而用多粘菌素B刺激分泌的肥大细胞则显示出强烈的荧光染料与单个细胞质颗粒结合。将活体黄连素染色后细胞群体的平均荧光强度与肝素和组胺释放进行了比较。结果强烈表明，在所使用的活体染色条件下，黄连素是已释放组胺的细胞内颗粒的标志物。活体染色方法还用于研究多粘菌素B诱导分泌后的膜事件。在多粘菌素B刺激终止后的第一个小时内，平均荧光强度下降了75%，而肥大细胞中的组胺和肝素含量保持不变。这些发现支持了这样一种观点，即肥大细胞分泌后膜会迅速恢复，允许选择性胺释放而不伴随肝素或颗粒的其他基质成分的释放。
