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通过细胞荧光测定法和高效液相色谱法研究大鼠肥大细胞中多巴胺的摄取和周转。

Uptake and turnover of dopamine in rat mast cells studied by cytofluorometry and high performance liquid chromatography.

作者信息

Rundquist I, Allenmark S, Enerbäck L

出版信息

Histochem J. 1982 May;14(3):429-43. doi: 10.1007/BF01011855.

Abstract

Uptake and turnover of dopamine (DA) in rat peritoneal mast cells were studied by a cytofluorometric technique. The main advantage of the method is that it permits the study of the distribution of amine content within populations of cells. Catecholamines and indolamines can be differentiated, but subtler structural differences in this group of compounds cannot be distinguished. We, therefore, combined the cytofluorometric measurements with a liquid chromatographic method based on reversed-phase chromatography followed by amperometric detection in a thin layer flow cell. Intraperitoneally injected L-DOPA was rapidly decarboxylated to DA, which was accumulated in mast cell granules. The elimination of DA from the mast cells was much faster than previously published 5-hydroxytryptamine and histamine elimination rates. No evidence of intracellular conversion of DA before its elimination was found and simultaneous heparin quantitations gave no evidence of an elimination pathway due to exocytosis of granules. Electron microscopy disclosed no structural changes that could be related to exocytosis during the elimination phase of DA. The rapid elimination together with absence of inhibition of DA-uptake after storage of exogenous 5-hydroxytryptamine suggest that the mechanism of DA storage differs from the mechanism of storage of endogenous mast cell amines.

摘要

采用细胞荧光测定技术研究了大鼠腹膜肥大细胞中多巴胺(DA)的摄取和周转。该方法的主要优点是能够研究细胞群体中胺含量的分布情况。儿茶酚胺和吲哚胺可以区分,但这组化合物中更细微的结构差异无法辨别。因此,我们将细胞荧光测定与一种基于反相色谱,随后在薄层流通池中进行安培检测的液相色谱方法相结合。腹腔注射的左旋多巴迅速脱羧转化为多巴胺,后者积聚在肥大细胞颗粒中。多巴胺从肥大细胞中的消除速度比之前报道的5-羟色胺和组胺的消除速度快得多。未发现多巴胺在消除前有细胞内转化的迹象,同时肝素定量分析也未发现因颗粒胞吐作用而产生的消除途径的证据。电子显微镜检查未发现多巴胺消除阶段与胞吐作用相关的结构变化。多巴胺的快速消除以及外源性5-羟色胺储存后多巴胺摄取未受抑制,这表明多巴胺的储存机制与内源性肥大细胞胺的储存机制不同。

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