title: Membrane-associated prostaglandin E synthase-1 is upregulated by proinflammatory cytokines in chondrocytes from patients with osteoarthritis.
abstract: Prostaglandin E synthase (PGES) including isoenzymes of membrane-associated PGES (mPGES)-1, mPGES-2, and cytosolic PGES (cPGES) is the recently identified terminal enzyme of the arachidonic acid cascade. PGES converts prostaglandin (PG)H2 to PGE2 downstream of cyclooxygenase (COX). We investigated the expression of PGES isoenzyme in articular chondrocytes from patients with osteoarthritis (OA). Chondrocytes were treated with various cytokines and the expression of PGES isoenzyme mRNA was analyzed by the reverse transcription-polymerase chain reaction and Northern blotting, whereas Western blotting was performed for protein expression. The subcellular localization of mPGES-1 was determined by immunofluorescent microscopy. Conversion of arachidonic acid or PGH2 to PGE2 was measured by enzyme-linked immunosorbent assay. Finally, the expression of mPGES-1 protein in OA articular cartilage was assessed by immunohistochemistry. Expression of mPGES-1 mRNA in chondrocytes was significantly induced by interleukin (IL)-1beta or tumor necrosis factor (TNF)-alpha, whereas other cytokines, such as IL-4, IL-6, IL-8, IL-10, and interferon-gamma, had no effect. COX-2 was also induced under the same conditions, although its pattern of expression was different. Expression of cPGES, mPGES-2, and COX-1 mRNA was not affected by IL-1beta or TNF-alpha. The subcellular localization of mPGES-1 and COX-2 almost overlapped in the perinuclear region. In comparison with 6-keto-PGF1alpha and thromboxane B2, the production of PGE2 was greater after chondrocytes were stimulated by IL-1beta or TNF-alpha. Conversion of PGH2 to PGE2 (PGES activity) was significantly increased in the lysate from IL-1beta-stimulated chondrocytes and it was inhibited by MK-886, which has an inhibitory effect on mPGES-1 activity. Chondrocytes in articular cartilage from patients with OA showed positive immunostaining for mPGES-1. These results suggest that mPGES-1 might be important in the pathogenesis of OA. It might also be a potential new target for therapeutic strategies that specifically modulate PGE2 synthesis in patients with OA.
authors.affiliation: Institute of Medical Science, St Marianna University School of Medicine, Kawasaki, Japan. kojimaf@marianna-u.ac.jp <kojimaf@marianna-u.ac.jp>
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Kojima
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Naraba
authors.forename: Satoshi
authors.initials: S
authors.lastname: Miyamoto
authors.forename: Moroe
authors.initials: M
authors.lastname: Beppu
authors.forename: Haruhito
authors.initials: H
authors.lastname: Aoki
authors.forename: Shinichi
authors.initials: S
authors.lastname: Kawai
mesh_terms: D015121:6-Ketoprostaglandin F1 alpha / Q000096:biosynthesis; D002358:Cartilage, Articular / Q000201:enzymology / Q000473:pathology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology / Q000502:physiology; D051546:Cyclooxygenase 2; D016207:Cytokines / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007249:Inflammation / Q000473:pathology; D007375:Interleukin-1 / Q000378:metabolism*; D019746:Intramolecular Oxidoreductases / Q000096:biosynthesis / Q000378:metabolism*; D007527:Isoenzymes / Q000096:biosynthesis; D008565:Membrane Proteins / Q000378:metabolism; D010003:Osteoarthritis / Q000201:enzymology* / Q000473:pathology; D010641:Phenotype; D000071876:Prostaglandin-E Synthases; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D013929:Thromboxane B2 / Q000096:biosynthesis; D013997:Time Factors; D015854:Up-Regulation / Q000502:physiology*; D002722:Chlorobenzenes
chemical_list: D015121:6-Ketoprostaglandin F1 alpha; D051546:Cyclooxygenase 2; D016207:Cytokines; D007211:Indoles; D007375:Interleukin-1; D019746:Intramolecular Oxidoreductases; D007527:Isoenzymes; D008565:Membrane Proteins; D000071876:Prostaglandin-E Synthases; D011451:Prostaglandin-Endoperoxide Synthases; D012333:RNA, Messenger; D013929:Thromboxane B2; C060893:MK-886; D002722:Chlorobenzenes; C496540:PTGS2 protein, human; C000605862:PTGES protein, human; C000605863:PTGES2 protein, human