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首页> 外文期刊>Life sciences >Caco-2 cells in culture synthesize and degrade dopamine and 5-hydroxytryptamine: A comparison with rat jejunal epithelial cells
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Caco-2 cells in culture synthesize and degrade dopamine and 5-hydroxytryptamine: A comparison with rat jejunal epithelial cells

机译：培养中的Caco-2细胞合成并降解多巴胺和5-羟色胺：与大鼠空肠上皮细胞的比较

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To explore the usefulness of Caco-2 cells in the study of intestinal dopaminergic and 5-hydroxytryptaminergic physiology, we have undertaken the study of aromatic L-amino acid decarboxylase (AADC), catechol-O-methyltransferase (COMT) and type A and B monoamine oxidase (MAO-A and MAO-B) activities in these cells using specific substrates. The activity of these enzymes was also evaluated in isolated rat jejunal epithelial cells. The results showed that V-max values (in nmol mg protein(-1) h(-1)) for AADC, using L-DOPA as the substrate, in rat jejunal epithelial cells (127.3+/-11.4) were found to be 6-fold higher than in Caco-2 cells (22.5 +/- 2.6). However, K-m values in Caco-2 cells (1.24+/-0.37 mM) were similar to those observed in rat jejuanl epithelial cells (1.30+/-0.29 mM). Similar results were obtained when AADC activity was evaluated using L-5HTP as substrate; in rat jejunal epithelial cells V-max values (in nmol mg prot(-1) h(-1)) were found to be 5-fold that in Caco-2 cells (16.3+/-1.0 and 3.0+/-0.2, respectively), and K-m values in Caco-2 cells (0.23+/-0.08 mM) were again similar to those observed in rat intestinal epithelial cells (0.09+/-0.03 mM). Caco-2 cells were not able to O-methylate dopamine, in contrast to rat jejunal epithelial cells (V-max = 8.6 +/- 0.4 nmol mg protein(-1) h(-1); K-m = 516+/-57 mu M). V-max values (in nmol mg protein(-1) h(-1)) for type A and B MAO in Caco-2 cells (19.0+/-0.6 and 5.4+/-0.6, respectively) were found to be significantly lower (P<0.05) than those in rat jejunal epithelial cells (46.9+/-3.1 and 9.6+/-1.2, respectively); however, no significant differences in the K-m values were observed between Caco-2 and rat jejunal epithelial cells for both type A and B MAO. In conclusion, Caco-2 cells in culture are endowed with the synthetic and metabolic machinery needled to form and degrade DA and 5-HT, though, no COMT activity could be detected in these cells. [References: 34]
机译：为了探索Caco-2细胞在肠道多巴胺能和5-羟色胺能生理学研究中的有用性，我们进行了芳香族L-氨基酸脱羧酶（AADC），儿茶酚-O-甲基转移酶（COMT）以及A型和B型研究单胺氧化酶（MAO-A和MAO-B）在这些细胞中使用特定的底物进行活性。还在分离的大鼠空肠上皮细胞中评估了这些酶的活性。结果显示，在大鼠空肠上皮细胞（127.3 +/- 11.4）中，以L-DOPA为底物，AADC的V-max值（nmol mg蛋白（-1）h（-1））为比Caco-2细胞高2倍（22.5 +/- 2.6）。但是，Caco-2细胞中的K-m值（1.24 +/- 0.37 mM）与大鼠空肠上皮细胞中观察到的K-m值（1.30 +/- 0.29 mM）相似。当使用L-5HTP作为底物评估AADC活性时，获得了相似的结果。在大鼠空肠上皮细胞中，V-max值（nmol mg prot（-1）h（-1））是Caco-2细胞的5倍（16.3 +/- 1.0和3.0 +/- 0.2， Caco-2细胞（0.23 +/- 0.08 mM）的Km值再次与大鼠肠上皮细胞（0.09 +/- 0.03 mM）的相似。与大鼠空肠上皮细胞相比，Caco-2细胞不能O-甲基化多巴胺（V-max = 8.6 +/- 0.4 nmol mg蛋白（-1）h（-1）; Km = 516 +/- 57亩M）。发现Caco-2细胞中A型和B型MAO的V-max值（以nmol mg蛋白（-1）h（-1）表示）（分别为19.0 +/- 0.6和5.4 +/- 0.6）显着低于大鼠空肠上皮细胞（分别为46.9 +/- 3.1和9.6 +/- 1.2）（P <0.05）;但是，对于A型和B型MAO，Caco-2和大鼠空肠上皮细胞之间的K-m值均未观察到显着差异。总之，培养的Caco-2细胞具有合成和代谢机制，可形成和降解DA和5-HT，尽管在这些细胞中未检测到COMT活性。 [参考：34]

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