Crooks P A, Breakefield X O, Sulens C H, Castiglione C M, Coward J K
Biochem J. 1978 Oct 15;176(1):187-96. doi: 10.1042/bj1760187.
[G-3H]Dopamine (3,4-dihydroxyphenethylamine) metabolism in human skin fibroblasts and rat hepatoma cells in culture was determined by high-pressure liquid-chromatographic analysis of both cell extract and uptake medium. Conjugated metabolites were selectively hydrolysed by incubation with arylsulphatase or beta-glucuronidase before analysis. The principal metabolites of dopamine in fibroblast cells are 3-methoxytyramine 4-O-sulphate and 3-methoxytyramine. No significant differences, either in the amounts of these metabolites or in the amount of dopamine metabolism, were observed in fibroblasts from both normal and homocystinuric individuals. In rat hepatoma cells, the major metabolite of dopamine was 3-methoxytyramine 4- or 3-O-glucuronide; lower concentrations of dopamine 4- or 3-O-glucuronide, 4-hydroxy-3-methoxyphenylacetic acid, 3,4-dihydroxyphenylacetic acid and two unidentified glucuronide conjugates were also observed. Significant differences in the relative concentrations of these metabolites in cell and uptake medium were observed in both cell systems.
通过对细胞提取物和摄取培养基进行高压液相色谱分析,测定了培养的人皮肤成纤维细胞和大鼠肝癌细胞中[G-3H]多巴胺(3,4-二羟基苯乙胺)的代谢情况。在分析之前,通过与芳基硫酸酯酶或β-葡萄糖醛酸酶孵育,选择性地水解结合代谢物。成纤维细胞中多巴胺的主要代谢物是3-甲氧基酪胺4-O-硫酸盐和3-甲氧基酪胺。在正常个体和同型胱氨酸尿症个体的成纤维细胞中,这些代谢物的量或多巴胺代谢量均未观察到显著差异。在大鼠肝癌细胞中,多巴胺的主要代谢物是3-甲氧基酪胺4-或3-O-葡萄糖醛酸苷;还观察到较低浓度的多巴胺4-或3-O-葡萄糖醛酸苷、4-羟基-3-甲氧基苯乙酸、3,4-二羟基苯乙酸和两种未鉴定的葡萄糖醛酸苷结合物。在这两种细胞系统中,均观察到这些代谢物在细胞和摄取培养基中的相对浓度存在显著差异。