Cytochrome P450 Metabolites of Arachidonic Acid in Human Placenta

Prostaglandins Pub Date : 1997-10-01 DOI:10.1016/S0090-6980(97)00148-2

W.R Schaefer , K Werner , H Schweer , J Schneider , E Arbogast , H.P Zahradnik

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引用次数: 16

Abstract

Little is known about the epoxygenase pathway of the arachidonic acid cascade in uterine tissues. In this paper, we describe the formation of epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETs) in human term placenta after uncomplicated pregnancies. Metabolism of [³H]-arachidonic acid was analyzed in short term tissue cultures of placenta by reverse phase HPLC. Major metabolites coeluted with authentic EETs and DHETs. The formation of EETs was inhibited by carbon monoxide. In non-radioactive incubations with biopsies from seven different placentas, sufficient material for GC/MS analysis was obtained. The combined media were purified by solid phase extraction and reverse phase HPLC. The fraction coeluting with DHETs was derivatized with pentafluorobenzylbromide (PFB) and bis-(trimethylsilyl)-trifluoroacetylacetamide (BSTFA) and analyzed by GC/NICI/MS/MS. 11,12-DHET and 14,15-DHET were identified by their mass spectra displaying specific fragments at m/z 149 and m/z 189, respectively. Our results suggest that the epoxygenase pathway is active in human term placenta.

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花生四烯酸在人胎盘中的细胞色素P450代谢物

对子宫组织中花生四烯酸级联的环氧化酶途径知之甚少。在本文中，我们描述了无并发症妊娠后人类足月胎盘中环氧二十碳三烯酸(EETs)和二羟基二十碳三烯酸(DHETs)的形成。用反相高效液相色谱法分析了[3H]-花生四烯酸在胎盘短期组织培养中的代谢。主要代谢物与正宗的eet和DHETs结合。EETs的形成受到一氧化碳的抑制。在7个不同胎盘活检的非放射性培养中，获得了足够的GC/MS分析材料。采用固相萃取法和反相高效液相色谱法对组合培养基进行纯化。用五氟苯溴(PFB)和双(三甲基硅基)-三氟乙酰乙酰胺(BSTFA)衍生化，采用GC/NICI/MS/MS分析。11,12- dhet和14,15- dhet的质谱分别在m/z 149和m/z 189处显示特异片段。我们的研究结果表明，环氧合酶途径在人足月胎盘中是活跃的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Prostaglandins

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