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Title: The effects of COX-metabolites on cyclooxygenase-2 induction in LPS-treated endothelial cells
Authors: Pravit Akarasereenont
Sirikul Chotewuttakorn
Kitirat Techatraisak
Athiwat Thaworn
Mahidol University
Keywords: Medicine
Issue Date: 1-Dec-2001
Citation: Journal of the Medical Association of Thailand. Vol.84, No.SUPPL. 3 (2001)
Abstract: Cyclooxygenase (COX) is the first enzyme in the pathway in which arachidonic acid is converted to PGs, also called COX-metabolites. COX exists as COX-1 and COX-2 isoforms. Each COX-metabolite has different characters and functions. The amounts of each COX-metabolite produced in cells are also different depending on cell type and mitogen stimulated cells. These were thought to be autoregulation among COX-metabolites. Here, we have investigated the effects of COX-metabolites, such as PGI2, PGE2, PGF2α and U44069, on the induction of COX-2 in human umbilical vein endothelial cells (HUVEC) treated with LPS (1 μg/ml). COX activity was measured by the production of 6-keto-PGF1α, PGE2, PGF2α and TXB2 in the presence of exogenous arachidonic acids (10 μM for 10 min) using enzyme immunoassay (EIA). COX-1 and COX-2 protein was measured by immunoblotting using specific antibody. PGI2, PGE2, PGF2α or U44069, did not affect on basal COX activity in untreated HUVEC (24 h incubation). Untreated HUVEC contained COX-1 protein but not COX-2 protein. When HUVEC were treated with LPS (1 μg/ml for 24 h), COX activity and COX-2 protein was increased in a dose dependent manner. The increased COX activity in LPS (1 μg/ml) treated HUVEC was inhibited with PGE2 (0.03, 0.3 or 3 μM), but not PGI2, PGF2α or U44069, in a dose dependent manner. Similary, COX-2 protein expression in LPS treated HUVEC was also inhibited with PGE2, but not PGI2, PGF2α or U44069, in a dose dependent manner. These results suggested that PGE2, but not PGI2, PGF2α or TXA2 is a key in feedback regulation of COX-metabolites produced in HUVEC.
ISSN: 01252208
Appears in Collections:Scopus 2001-2005

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