LIPOXIN GENERATION BY HUMAN MEGAKARYOCYTE-INDUCED 12-LIPOXYGENASE
LIPOXIN GENERATION BY HUMAN MEGAKARYOCYTE-INDUCED 12-LIPOXYGENASE
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DOI：
10.1016/0167-4889(92)90073-k
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发表时间：
1992-01-13
期刊：
影响因子：
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通讯作者：
SERHAN, CN
中科院分区：
文献类型：
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作者：
SHEPPARD, KA;GREENBERG, SM;SERHAN, CN
用人巨核细胞系（Dami）检查类花生酸生物合成。与[1-C-14]花生四烯酸和离子载体A23187或凝血酶孵育的巨核细胞产生血栓烷和12-羟基十七碳三烯酸（HHTrE）。暴露于佛波醇肉豆蔻酸酯乙酸酯（PMA）1至9天诱导分化，并显示增加的[1-C-14]花生四烯酸转化为环氧合酶和脂氧合酶（LO）衍生的产品。通过GC/MS和手性柱SP-HPLC等物理特性鉴定了LO衍生产物为12 S-HETE。PMA处理的Dami细胞不产生5-HETE，白三烯或脂氧素从外源性花生四烯酸，而它们确实将白三烯A4（LTA 4）转化为脂氧素A4，脂氧素B4和它们各自的全反式异构体。此外，用人12-脂氧合酶cDNA转染的COS-M6细胞与花生四烯酸或LTA 4孵育分别产生12-HETE和脂氧素。与LTA 4孵育的转染的COS-M6细胞产生的脂氧素谱与PMA处理的Dami细胞产生的脂氧素谱相似。结果表明，人巨核细胞可以将花生四烯酸和LTA 4转化为具有生物活性的类花生酸，并且在这些细胞进一步分化后出现12-脂氧合酶。此外，他们表明人类巨核细胞的12-LO和转染的COS细胞表达的12-LO可以产生脂氧素A4和B4。它们一起表明，人12-LO可以作为LTA 4的LX-合成酶活性的模型。
Eicosanoid biosynthesis was examined with a human megakaryocytic cell line (Dami). Megakaryocytes incubated with [1-C-14]arachidonic acid and either ionophore A23187 or thrombin generated both thromboxane and 12-hydroxyheptadecatrienoic acid (HHTrE). Exposure to phorbol myristate acetate (PMA) for 1 through 9 days induced differentiation and revealed an increase in the conversion of [1-C-14]arachidonate to cyclooxygenase- and lipoxygenase (LO)-derived products. The LO-derived product was identified as 12S-HETE by its physical characteristics including GC/MS and chiral column SP-HPLC. PMA-treated Dami cells did not generate 5-HETE, leukotrienes or lipoxins from exogenous arachidonic acid while they did convert leukotriene A4 (LTA4) to lipoxin A4, lipoxin B4 and their respective all-trans isomers. In addition, COS-M6 cells transfected with a human 12-lipoxygenase cDNA and incubated with either arachidonic acid or LTA4 generated 12-HETE and lipoxins, respectively. The lipoxin profile generated by transfected COS-M6 cells incubated with LTA4 was similar to that generated by the PMA-treated Dami cells. Results indicate that human megakaryocytes can transform arachidonate and LTA4 to bioactive eicosanoids and that the 12-lipoxygenase appears upon further differentiation of these cells. In addition, they indicate that the 12-LO of human megakaryocytes and the 12-LO expressed by transfected COS cells can generate both lipoxins A4 and B4. Together they suggest that the human 12-LO can serve as a model of LX-synthetase activity with LTA4.