The effects of COX-metabolites on cyclooxygenase-2 induction in LPS-treated endothelial cells.
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, PGF2alpha and U44069, on the induction of COX-2 in human umbilical vein endothelial cells (HUVEC) treated with LPS (1 microg/ml). COX activity was measured by the production of 6-keto-PGF1alpha, PGE2, PGF2alpha and TXB2 in the presence of exogenous arachidonic acids (10 microM for 10 min) using enzyme immunoassay (EIA). COX-1 and COX-2 protein was measured by immunoblotting using specific antibody. PGI2, PGE2, PGF2alpha 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 microg/ml for 24 h), COX activity and COX-2 protein was increased in a dose dependent manner. The increased COX activity in LPS (1 microg/ml) treated HUVEC was inhibited with PGE2 (0.03, 0.3 or 3 microM), but not PGI2, PGF2alpha or U44069, in a dose dependent manner. Similarly, COX-2 protein expression in LPS treated HUVEC was also inhibited with PGE2, but not PG2, PGF2alpha or U44069, in a dose dependent manner. These results suggested that PGE2, but not PGI2, PGF2alpha or TXA2 is a key in feedback regulation of COX-metabolites produced in HUVEC.
Akarasereenont
P
P
Department of Pharmacology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Chotewuttakorn
S
S
Techatraisak
K
K
Thaworn
A
A
D002478:Cells, Cultured; D051546:Cyclooxygenase 2; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008565:Membrane Proteins; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D014471:Umbilical Veins / Q000166:cytology
D007527:Isoenzymes; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases