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Biosynthesis of lipoxygenase and cyclo-oxygenase products from [14C]-arachidonic acid by human colonic mucosa.
  1. N K Boughton-Smith,
  2. C J Hawkey,
  3. B J Whittle


    The human colon synthetises several prostanoids which may have a role in inflammatory bowel diseases. As lipoxygenase products of arachidonate metabolism have been implicated in inflammatory processes, we have now investigated the formation of both lipoxygenase and cyclo-oxygenase metabolites from [14C]-arachidonic acid [(14C]-AA) by human colonic tissue. Homogenates of human colonic mucosa were incubated with [14C]-AA and after extraction into diethyl ether, separated by thin layer chromatography using two solvent systems that allowed resolution of cyclo-oxygenase and lipoxygenase products. The predominant cyclo-oxygenase products, as identified by their chromatographic mobility, were PGE2 greater than PGF2 alpha greater than PGD2 greater than TXB2 greater than 6-keto-PGF1 alpha. The formation of these products was inhibited both by indomethacin (1-10 microM) and the dual pathway inhibitor, BW755C (1-30 microM). The predominant lipoxygenase products formed, which had the chromatographic mobility of 11-, 12-, 15-HETE (which ran together) were inhibited by BW755C (19 microM) but not by indomethacin (3 microM). Further resolution of this TLC band, performed using normal phase HPLC, indicated that both 12-HETE and 15-HETE were major lipoxygenase products formed by human colonic homogenate. The present findings indicate that human colonic tissue can convert [14C]-AA into lipoxygenase as well as cyclo-oxygenase products and support the suggestion that lipoxygenase products may have a role in inflammatory bowel disease.

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