Tragni E, Caruso D, Porta S, Fumagalli R, Galli G, Galli C L
Institute of Pharmacological Sciences, University of Milan, Italy.
Arch Dermatol Res. 1988;280(7):437-42. doi: 10.1007/BF00429984.
The established mouse epidermis-derived cell line HEL/30 was incubated in the presence of 3H arachidonic acid (AA) for 1 h. After medium removal, cells were reincubated with fresh medium in the presence or absence of the calcium ionophore A23187 and tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The AA metabolites formed were extracted from cell-free medium and analyzed using TLC and HPLC. The distribution of the recovered radioactivity showed PGE2, 15-hydroxy-eicosatetraenoic acid (15-HETE), and leukotriene B4 (LTB4), as major products of AA metabolism. The presence of calcium ionophore A23187 increased the release of radioactivity, without affecting the profile of metabolites present in the medium. TPA elicited a preferential increase of cycloxygenase metabolism, this effect being reversed by indomethacin. 5,8,11,14-eicosatetraynoic acid (ETYA) almost completely inhibited LT and HETE formation in A23187 and TPA-treated cells. The results show that HEL/30 cells are able to metabolize AA via both cyclo- and lipoxygenase pathways and that these activities can be modified by chemical means. This cell line might be a suitable tool for studying the involvement of arachidonic acid cascade in cell response to exogenous stimuli.
将已建立的小鼠表皮来源细胞系HEL/30在含有3H花生四烯酸（AA）的条件下孵育1小时。去除培养基后，将细胞在存在或不存在钙离子载体A23187和肿瘤启动子12-O-十四烷酰佛波醇-13-乙酸酯（TPA）的情况下，用新鲜培养基重新孵育。从无细胞培养基中提取形成的AA代谢产物，并使用薄层层析（TLC）和高效液相色谱（HPLC）进行分析。回收放射性的分布显示，前列腺素E2（PGE2）、15-羟基二十碳四烯酸（15-HETE）和白三烯B4（LTB4）是AA代谢的主要产物。钙离子载体A23187的存在增加了放射性的释放，但不影响培养基中代谢产物的谱。TPA引起环氧化酶代谢的优先增加，吲哚美辛可逆转此效应。5,8,11,14-二十碳四炔酸（ETYA）几乎完全抑制了A23187和TPA处理细胞中白三烯（LT）和HETE的形成。结果表明，HEL/30细胞能够通过环氧化酶和脂氧化酶途径代谢AA，并且这些活性可以通过化学方法进行调节。该细胞系可能是研究花生四烯酸级联反应在细胞对外源刺激反应中的作用的合适工具。
