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Microsomal prostaglandin E synthase‐1, which is not coupled to a particular cyclooxygenase isoenzyme, is essential for prostaglandin E 2 biosynthesis in vascular smooth muscle cells
Author(s) -
CAMACHO M.,
GERBOLÉS E.,
ESCUDERO J.R.,
ANTÓN R.,
GARCÍAMOLL X.,
VILA L.
Publication year - 2007
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/j.1538-7836.2007.02555.x
Subject(s) - cyclooxygenase , biosynthesis , prostaglandin , vascular smooth muscle , arachidonic acid , enzyme , prostaglandin e , prostaglandin h2 , prostaglandin e2 , cytosol , microsome , biochemistry , chemistry , incubation , microbiology and biotechnology , biology , endocrinology , smooth muscle
Summary. Background: Prostaglandin (PG) E 2 induces expression of matrix metalloproteinases and angiogenic factors, thereby contributing to plaque instability. Objective: To study the influence of cyclooxygenase (COX) and PGE synthase (PGES) isoenzyme expression on PGE 2 and PGI 2 biosynthesis in vascular smooth muscle cells (VSMC) in culture. Methods: Cells were treated with human recombinant IL‐1β over different periods of time. Expression of PGI synthase, and COX and PGES isoenzymes was determined by real‐time reverse transcriptase polymerase chain reaction and immunoblotting. Biosynthesis of prostanoids from exogenous or endogenous substrate was analyzed by high‐performance liquid chromatography or enzyme‐immunoassay after incubation of cells with labeled arachidonic acid or thrombin, respectively. Results: Cytosolic PGES and microsomal PGES (mPGES) ‐1 and ‐2 were expressed in VSMC. PGES activity was mainly linked to mPGES‐1. IL‐1β induced COX‐2 and mPGES‐1 with a different time course. VSMC ability to synthesize PGE 2 and PGI 2 fitted mPGES‐1 and COX‐2 expression, respectively. The ability of VSMC to produce PGI 2 was downregulated by mPGES‐1 expression and was restored when mPGES‐1 expression was silenced. Results from COX‐1 and COX‐2 silencing and selective inhibition showed that both COX‐1 and COX‐2 were involved in the biosynthesis of PGE 2 and their relative contribution depended on the time of incubation with IL‐1β. Conclusions: mPGES‐1 is the main PGES responsible for PGE 2 biosynthesis by VSMC and its induction downregulates VSMC ability to produce PGI 2. These results support the concept that under inflammatory conditions VSMC could significantly contribute to plaque instability and that mPGES‐1 may be a target for therapeutic intervention in patients with cardiovascular risk.