DOCSLIB.ORG

Expression and Regulation of Microsomal Prostaglandin E Synthase

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Expression and Regulation of Microsomal Prostaglandin E Synthase Expression and Regulation of Microsomal Prostaglandin E Synthase-1 in Human Osteoarthritic Cartilage and Chondrocytes XINFANG LI, HASSAN AFIF, SARANETTE CHENG, JOHANNE MARTEL-PELLETIER, JEAN-PIERRE PELLETIER, PIERRE RANGER, and HASSAN FAHMI ABSTRACT. Objective. Elevated production of prostaglandin E2 (PGE2) plays an important role in the pathogen- esis of arthritis. Recently, an inducible microsomal prostaglandin E synthase-1 (mPGES-1) was identified. This enzyme is functionally coupled with cyclooxygenase-2 (COX-2) and converts the COX product PGH2 to PGE2. We analyzed expression of mPGES-1 in human normal and osteoarthritic (OA) cartilage and determined the effect of different inflammatory agonists on the expression of mPGES-1 in OA chondrocytes. Methods. Expression of mPGES-1 mRNA and protein in cartilage was determined by quantitative real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry, respectively. OA chondrocytes were treated with different inflammatory agents, and mPGES-1 protein expression was evaluated by Western blot. Activation of the mPGES-1 promoter was assessed in transient trans- fection experiments. Results. Levels of mPGES-1 mRNA and protein were markedly elevated in OA versus normal car- tilage. Treatment of chondrocytes with interleukin 1ß (IL-1ß) induced expression of mPGES-1 pro- tein in a dose- and time-dependent manner. This appears to occur at the transcriptional level, as IL- 1ß induced expression of mPGES-1 mRNA and the activity of this gene promoter. Tumor necrosis factor-α (TNF-α) and IL-17 also upregulated expression of mPGES-1 protein and displayed a syn- ergistic effect with IL-1ß. Peroxisome proliferator-activated receptor-γ ligands, 15-deoxy-∆12,14- prostaglandin J2 and troglitazone, inhibited IL-1ß-induced mPGES-1 protein expression, an effect that was reversed by exogenous PGE2. Conclusion. Our study shows that mPGES-1 expression is upregulated in OA versus normal carti- lage and that proinflammatory cytokines increased mPGES-1 expression in chondrocytes. These data suggest that mPGES-1 may prove to be an interesting therapeutic target for controlling PGE2 synthesis. (J Rheumatol 2005;32:887–95) Key Indexing Terms: MICROSOMAL PROSTAGLANDIN E SYNTHASE-1 CARTILAGE CHONDROCYTES OSTEOARTHRITIS 1 Biochemical, genetic, and clinical evidence indicates that of PGE2 . Treatment with neutralizing anti-PGE2 antibodies prostaglandin E2 (PGE2) plays a critical role in inflamma- prevents acute and chronic inflammation in a rat adjuvant 2 tion and in the pathophysiology of articular joint diseases, arthritis model . More direct evidence for the role of PGE2 such as rheumatoid arthritis (RA) and osteoarthritis (OA). in arthritis has been provided by gene targeting studies. 3 For example, arthritic joint tissues produce large quantities Genetic disruption of either the PGE2 receptor EP4 or cyclooxygenase-2 (COX-2)4, one of the key enzymes in From the Osteoarthritis Research Unit, Centre hospitalier de l’Université PGE2 biosynthesis, reduced incidence and severity of colla- de Montréal, Hôpital Notre-Dame, Montréal, Québec, Canada. gen-induced arthritis in mice. These animals showed Supported by the Canadian Institutes of Health Research (CIHR), Grant IMH-63168, and the Fonds de Recherche en santé du Québec (FRSQ) reduced inflammation and less cartilage and bone destruc- Subvention d’Établissement de Jeune Chercheur JC2836. S. Cheng is tion. The role of PGE2 in arthritis is also supported by effec- supported by a fellowship from the CIHR Training on Mobility and Posture tive suppression of pain and inflammatory responses in Deficiencies (MENTOR). H. Fahmi is a Research Scholar of FRSQ. arthritis by nonsteroidal antiinflammatory drugs (NSAID) X. Li, MSc; H. Afif, PhD; S. Cheng, MSc; J. Martel-Pelletier, PhD; 5,6 J-P Pelletier, MD; H. Fahmi, PhD, Osteoarthritis Research Unit, Centre that reduce PGE2 biosynthesis . hospitalier de l’Université de Montréal; P. Ranger, MD, Hopital Sacré- Chondrocytes are a major source of PGE2 in the joint; the Coeur. production of this prostanoid can be induced by proinflam- Address reprint requests to H. Fahmi, Osteoarthritis Research Unit, matory cytokines, mitogens, mechanical stress, and trau- Centre hospitalier de l’Université de Montréal, Hôpital Notre-Dame, 5,7,8 1560 rue Sherbrooke est, Montréal, Québec H2L 4M1, Canada. ma . The synthesis of PGE2 from arachidonic acid (AA) E-mail: [email protected] requires 2 enzymes acting sequentially. Cyclooxygenases Accepted for publication December 29, 2004. catalyze the conversion of AA to the intermediate prostanoid Personal non-commercial use only. The Journal of Rheumatology Copyright © 2005. All rights reserved. Li, et al: PGE synthase in OA cartilage 887 Downloaded on September 27, 2021 from www.jrheum.org 16 PGH2. Two isoforms of the COX enzyme have been identi- digestion as described . Briefly, this consisted of 2 mg/ml pronase for 1 h fied: COX-1 is constitutively expressed in most tissues, followed by 1 mg/ml collagenase for 6 h (type IV; Sigma-Aldrich) at 37°C whereas COX-2 is induced by various stimuli including in DMEM and antibiotics (100 U/ml penicillin, 100 µg/ml streptomycin). The digested tissue was briefly centrifuged and the pellet was washed. The lipopolysaccharides, growth factors, and proinflammatory isolated chondrocytes were seeded at high density in tissue culture flasks 5,9 cytokines (reviewed in ). Subsequently, PGE synthase and cultured in DMEM supplemented with 10% heat-inactivated FCS. At (PGES) converts COX-derived PGH2 into PGE2. At least 3 confluence, the chondrocytes were detached, seeded at high density, and distinct PGES isoforms have been identified10. Cytosolic allowed to grow in DMEM, supplemented as above. The culture medium PGES (cPGES), which is identical to the heat shock protein was changed every second day, and 24 h before the experiment the cells were incubated in fresh medium containing 0.5% FCS. Only first-passage 90-associated protein p23, is ubiquitously and constitutive- chondrocytes were used. ly expressed, and displays functional coupling with COX-1. Immunohistochemistry. Cartilage specimens were processed for immuno- In contrast, microsomal PGES-1 (mPGES-1), originally histochemistry as described16. Specimens were fixed in 4% paraformalde- designated microsomal glutathione S-transferase 1-like 1, is hyde and embedded in paraffin. Sections (5 µm) of paraffin-embedded an inducible enzyme that exhibits preferential functional specimens were deparaffinized in toluene, and dehydrated in a graded coupling with COX-2. The most recently identified isoform, series of ethanol. The specimens were then preincubated with chondroiti- nase ABC (0.25 U/ml in phosphate buffered saline, PBS, pH 8.0) for 60 mPGES-2, is ubiquitously expressed in diverse tissues, but min at 37°C, followed by 30 min incubation with Triton X-100 (0.3%) at 10 its function and regulation remain obscure . room temperature. Slides were then washed in PBS followed by 2% hydro- The upregulation of mPGES-1 expression has been gen peroxide/methanol for 15 min. They were further incubated for 60 min with 2% normal serum (Vector Laboratories, Burlingame, CA, USA) and reported in conditions in which PGE2 has been implicated, such as arthritis11, and studies with mPGES-1-deficient overlaid with primary antibody for 18 h at 4°C in a humidified chamber. The antibody was a rabbit polyclonal anti-human mPGES-1 (Cayman), mice have shown that induced PGE2 synthesis is largely used at 10 µg/ml. Each slide was washed 3 times in PBS pH 7.4 and stained 12,13 dependent on this enzyme . Proinflammatory cytokines using the avidin-biotin complex method (Vectastain ABC kit; Vector). The interleukin 1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) color was developed with 3,3’-diaminobenzidine (DAB; Vector) containing have been shown to induce mPGES-1 expression in several hydrogen peroxide. Slides were counterstained with eosin. The specificity tissues and cell types, including synovial fibroblasts and of staining was evaluated by using antibody that had been preadsorbed (1 14,15 h, 37°C) with a 20-fold molar excess of the specific corresponding peptide, osteoblasts . However, little is known about the expres- and by substituting the primary antibody with non-immune rabbit IgG sion and regulation of mPGES-1 in cartilage. (Chemicon, Temecula, CA, USA; used at the same concentration as the To understand the regulation of PGE2 production in joint primary antibodies). The evaluation of positive-staining chondrocytes was tissues, we analyzed mPGES-1 expression in normal and performed using our published method16. For each specimen, 6 microscop- OA cartilage. Further, we explored the effect of different ic fields were examined under 40× magnification. The total number of chondrocytes and the number of chondrocytes staining positive were eval- inflammatory agonists on the expression of mPGES-1 in uated and results were expressed as the percentage of chondrocytes stain- OA chondrocytes. ing positive (cell score). RNA extraction and reverse transcriptase-polymerase chain reaction (RT- MATERIALS AND METHODS PCR). Total RNA from homogenized cartilage or stimulated chondrocytes Reagents. Recombinant human (rh) IL-1ß was obtained from Genzyme was isolated using the TRIzol® reagent (Invitrogen) according to the man- (Cambridge, MA, USA), rhTNF-α and rhIL-17 were from R&D Systems ufacturer’s instructions. To remove contaminating DNA, isolated RNA was ∆12,14 (Minneapolis, MN, USA). 15-deoxy- -prostaglandin J2 (15d-PGJ2),
Load more

Recommended publications
  • Prostaglandin E2 Deficiency Uncovers a Dominant Role for Thromboxane A2
    Prostaglandin E2 deficiency uncovers a dominant role for thromboxane A2 in house dust mite-induced allergic pulmonary inflammation Tao Liua,b, Tanya M. Laidlawa,b,c, Chunli Fenga,b, Wei Xinga,b, Shiliang Shena,b, Ginger L. Milned, and Joshua A. Boycea,b,c,1 Departments of aMedicine and cPediatrics, Harvard Medical School, Boston, MA 02115; bDivision of Rheumatology, Immunology, and Allergy, Jeff and Penny Vinik Center for Allergic Disease Research, Brigham and Women’s Hospital, Boston, MA 02115; and dDepartment of Pharmacology, Vanderbilt University, Nashville, TN 37232 Edited* by K. Frank Austen, Brigham and Women’s Hospital, Boston, MA, and approved June 20, 2012 (received for review May 10, 2012) fl – Prostaglandin E2 (PGE2) is an abundant lipid in ammatory media- PGE2 production (16 19) and reduced expression of the EP2 re- tor with potent but incompletely understood anti-inflammatory ceptor (20, 21), as well as marked tissue eosinophilia and bron- actions in the lung. Deficient PGE2 generation in the lung predis- choconstrictive responses to the administration of nonselective poses to airway hyperresponsiveness and aspirin intolerance in COX inhibitors. These findings suggest potential therapeutic fi −/− asthmatic individuals. PGE2-de cient ptges mice develop exag- applications of PGE2 in asthma and AERD if the mechanisms gerated pulmonary eosinophilia and pulmonary arteriolar smooth- responsible for the homeostatic functions of PGE2 in asthma can fi muscle hyperplasia compared with PGE2-suf cient controls when be fully defined. challenged intranasally with a house dust mite extract. We now An extract (Df) of the house dust mite Dermatophagoides demonstrate that both pulmonary eosinophilia and vascular farinae contains clinically relevant protease allergens, as well as fi remodeling in the setting of PGE2 de ciency depend on thrombox- adjuvants (glycans, endotoxin) that elicit sensitization through ane A2 and signaling through the T prostanoid (TP) receptor.
    [Show full text]
  • Coexpression of Microsomal Prostaglandin E Synthase With
    Coexpression of Microsomal Prostaglandin E Synthase with Cyclooxygenase-2 in Human Rheumatoid Synovial Cells FUMIAKI KOJIMA, HIROAKI NARABA, YASUHARU SASAKI, RENZO OKAMOTO, TOMIHISA KOSHINO, and SHINICHI KAWAI ABSTRACT. Objective. Recently, microsomal prostaglandin (PG) E synthase (mPGES) was cloned as a terminal enzyme catalyzing PGH2 to PGE2. We investigated mPGES as well as cyclooxygenase (COX)-2, catalyzing arachidonic acid to PGH2, in synovial cells from patients with rheumatoid arthritis (RA). The effect of dexamethasone on mPGES expression was also studied. Methods. Synovial cells were treated with interleukin 1ß (IL-1ß) and dexamethasone under various conditions, and expression of mPGES mRNA and protein was analyzed by Northern blot and Western blot, respectively. Conversions of arachidonic acid or PGH2 to PGE2 were measured by ELISA. Subcellular localization of mPGES and COX-2 was determined by immunofluorescent microscopic analysis. Results. mPGES mRNA and protein expression were significantly upregulated by IL-1ß in synovial cells. COX-2 mRNA and protein were also upregulated by IL-1ß, but with a different time course from that of mPGES. Conversion of PGH2 to PGE2 was increased by IL-1ß and was correlated with mPGES expression. Increased conversion of arachidonic acid to PGE2 was maintained when mPGES and COX-2 were coexpressed. Subcellular localization of mPGES and COX-2 overlapped in the perinuclear region in IL-1ß stimulated synovial cells. Dexamethasone inhibited mRNA and protein expression for mPGES and increased conversion of arachidonic acid to PGE2, but inhibition of mPGES was weaker compared with that of COX-2 in IL-1ß stimulated cells. Conclusion. The results suggest that abundant PGE2 production at inflammation sites such as rheumatoid synovia is caused by the coordinated upregulation of mPGES and COX-2.
    [Show full text]
  • Effect of Prostanoids on Human Platelet Function: an Overview
    International Journal of Molecular Sciences Review Effect of Prostanoids on Human Platelet Function: An Overview Steffen Braune, Jan-Heiner Küpper and Friedrich Jung * Institute of Biotechnology, Molecular Cell Biology, Brandenburg University of Technology, 01968 Senftenberg, Germany; steff[email protected] (S.B.); [email protected] (J.-H.K.) * Correspondence: [email protected] Received: 23 October 2020; Accepted: 23 November 2020; Published: 27 November 2020 Abstract: Prostanoids are bioactive lipid mediators and take part in many physiological and pathophysiological processes in practically every organ, tissue and cell, including the vascular, renal, gastrointestinal and reproductive systems. In this review, we focus on their influence on platelets, which are key elements in thrombosis and hemostasis. The function of platelets is influenced by mediators in the blood and the vascular wall. Activated platelets aggregate and release bioactive substances, thereby activating further neighbored platelets, which finally can lead to the formation of thrombi. Prostanoids regulate the function of blood platelets by both activating or inhibiting and so are involved in hemostasis. Each prostanoid has a unique activity profile and, thus, a specific profile of action. This article reviews the effects of the following prostanoids: prostaglandin-D2 (PGD2), prostaglandin-E1, -E2 and E3 (PGE1, PGE2, PGE3), prostaglandin F2α (PGF2α), prostacyclin (PGI2) and thromboxane-A2 (TXA2) on platelet activation and aggregation via their respective receptors. Keywords: prostacyclin; thromboxane; prostaglandin; platelets 1. Introduction Hemostasis is a complex process that requires the interplay of multiple physiological pathways. Cellular and molecular mechanisms interact to stop bleedings of injured blood vessels or to seal denuded sub-endothelium with localized clot formation (Figure1).
    [Show full text]
  • Prostaglandin Signaling Regulates Nephron Segment Patterning Of
    RESEARCH ARTICLE Prostaglandin signaling regulates nephron segment patterning of renal progenitors during zebrafish kidney development Shahram Jevin Poureetezadi1,2, Christina N Cheng1,2, Joseph M Chambers1,2, Bridgette E Drummond1,2, Rebecca A Wingert1,2* 1Department of Biological Sciences, University of Notre Dame, Notre Dame, United States; 2Center for Stem Cells and Regenerative Medicine, Center for Zebrafish Research, University of Notre Dame, Notre Dame, United States Abstract Kidney formation involves patterning events that induce renal progenitors to form nephrons with an intricate composition of multiple segments. Here, we performed a chemical genetic screen using zebrafish and discovered that prostaglandins, lipid mediators involved in many physiological functions, influenced pronephros segmentation. Modulating levels of prostaglandin E2 (PGE2) or PGB2 restricted distal segment formation and expanded a proximal segment lineage. Perturbation of prostaglandin synthesis by manipulating Cox1 or Cox2 activity altered distal segment formation and was rescued by exogenous PGE2. Disruption of the PGE2 receptors Ptger2a and Ptger4a similarly affected the distal segments. Further, changes in Cox activity or irx3b sim1a PGE2 levels affected expression of the transcription factors and that mitigate pronephros segment patterning. These findings show for the first time that PGE2 is a regulator of nephron formation in the zebrafish embryonic kidney, thus revealing that prostaglandin signaling may have implications for renal birth defects and other diseases. DOI: 10.7554/eLife.17551.001 *For correspondence: rwingert@ nd.edu Introduction Competing interests: The The kidney serves central functions in metabolic waste excretion, osmoregulation, and electrolyte authors declare that no homeostasis. Vertebrate kidney organogenesis is a dynamic process involving the generation of up competing interests exist.
    [Show full text]
  • Figure S1. HAEC ROS Production and ML090 NOX5-Inhibition
    Figure S1. HAEC ROS production and ML090 NOX5-inhibition. (a) Extracellular H2O2 production in HAEC treated with ML090 at different concentrations and 24 h after being infected with GFP and NOX5-β adenoviruses (MOI 100). **p< 0.01, and ****p< 0.0001 vs control NOX5-β-infected cells (ML090, 0 nM). Results expressed as mean ± SEM. Fold increase vs GFP-infected cells with 0 nM of ML090. n= 6. (b) NOX5-β overexpression and DHE oxidation in HAEC. Representative images from three experiments are shown. Intracellular superoxide anion production of HAEC 24 h after infection with GFP and NOX5-β adenoviruses at different MOIs treated or not with ML090 (10 nM). MOI: Multiplicity of infection. Figure S2. Ontology analysis of HAEC infected with NOX5-β. Ontology analysis shows that the response to unfolded protein is the most relevant. Figure S3. UPR mRNA expression in heart of infarcted transgenic mice. n= 12-13. Results expressed as mean ± SEM. Table S1: Altered gene expression due to NOX5-β expression at 12 h (bold, highlighted in yellow). N12hvsG12h N18hvsG18h N24hvsG24h GeneName GeneDescription TranscriptID logFC p-value logFC p-value logFC p-value family with sequence similarity NM_052966 1.45 1.20E-17 2.44 3.27E-19 2.96 6.24E-21 FAM129A 129. member A DnaJ (Hsp40) homolog. NM_001130182 2.19 9.83E-20 2.94 2.90E-19 3.01 1.68E-19 DNAJA4 subfamily A. member 4 phorbol-12-myristate-13-acetate- NM_021127 0.93 1.84E-12 2.41 1.32E-17 2.69 1.43E-18 PMAIP1 induced protein 1 E2F7 E2F transcription factor 7 NM_203394 0.71 8.35E-11 2.20 2.21E-17 2.48 1.84E-18 DnaJ (Hsp40) homolog.
    [Show full text]
  • Supplementary Table 2
    Supplemental Table 2. Genes differentially expressed between Oct-1 deficient and wild-type primary MEFs in response to 10 Gy gamma radiation Fold Fold Fold Fold Symbol; Gene title* Probeset† Accession WTR/WT AdjP-value MUR/MU AdjP-value MUR/WTR AdjP-value MU/WT AdjP-value Rank WT,WTR,MU,MUR‡ Stress/oxidative response Sesn2; sestrin 2 1425139_at AV308638 1.96 2.29E-03 1.63 0.27 1.04 0.99 1.25 0.99 9433,7042,8664,6933 Sesn2; sestrin 2 1451599_at AV308638 1.89 1.75E-02 1.37 0.99 -1.12 0.99 1.23 0.99 10113,7936,9436,8378 Sdfr2; stromal cell derived factor receptor 2 1423465_at BB032852 1.79 4.86E-02 1.09 0.99 -1.75 5.89E-02 -1.06 0.99 12658,11138,12863,12615 Ddit4; DNA-damage-inducible transcript 4 1428306_at AK017926 1.73 8.71E-03 1.48 0.52 1.11 0.99 1.30 0.59 4972,3301,4100,3098 Bbc3; Bcl-2 binding component 3 1423315_at AW489168 1.72 2.85E-02 1.44 0.96 -1.18 0.99 1.01 0.99 8741,6780,8712,7427 Icam1; intercellular adhesion molecule-1 1424067_at BC008626 1.64 8.42E-05 6.24 <1E-9 4.11 <1E-9 1.08 0.99 19383,16809,19072,12583 Homer3; homer homolog 3 1424859_at BC005773 1.57 0.26 1.76 1.40E-02 1.10 0.99 -1.03 0.99 8645,7023,8730,6719 Daf1; decay accelerating factor 1 1443906_at BE686894 1.54 3.25E-02 1.50 0.99 1.69 1.45E-02 1.73 3.48E-04 7091,5401,4959,3865 Prkaa2; protein kinase, AMP-activated, alpha 2 catalytic subunit 1434766_at BQ175911 1.48 6.67E-08 1.09 0.99 -1.37 2.34E-05 -1.00 0.99 14476,10813,14580,13900 Tnfrsf10b; tumor necrosis factor receptor superfamily, member 10b 1421296_at NM_020275 1.46 0.99 2.03 4.75E-03 -1.21 0.99 -1.68 7.78E-03 14962,13936,17023,14476 Ccng1; cyclin G1 1420827_a_at BG065754 1.45 3.60E-03 1.23 0.99 -1.07 0.99 1.10 0.99 786,528,726,576 Prkaa2; protein kinase, AMP-activated, alpha 2 catalytic subunit 1429463_at BB612385 1.42 <1E-9 1.10 0.24 -1.30 <1E-9 1.00 0.99 21376,18687,21380,20908 Nxn; nucleoredoxin 1422465_a_at BB366804 1.29 0.76 1.42 4.63E-02 1.17 0.99 1.06 0.99 2272,1839,2180,1619 Cox10; cytochrome c oxidase assemb.
    [Show full text]
  • Synthase-2 2 Stem Cells by Targeting Prostaglandin E Immunoregulatory
    MicroRNA-146a Negatively Regulates the Immunoregulatory Activity of Bone Marrow Stem Cells by Targeting Prostaglandin E2 Synthase-2 This information is current as of September 30, 2021. Mariola Matysiak, Maria Fortak-Michalska, Bozena Szymanska, Wojciech Orlowski, Anna Jurewicz and Krzysztof Selmaj J Immunol 2013; 190:5102-5109; Prepublished online 15 April 2013; Downloaded from doi: 10.4049/jimmunol.1202397 http://www.jimmunol.org/content/190/10/5102 http://www.jimmunol.org/ References This article cites 45 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/190/10/5102.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 30, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology MicroRNA-146a Negatively Regulates the Immunoregulatory Activity of Bone Marrow Stem Cells by Targeting Prostaglandin E2 Synthase-2 Mariola Matysiak, Maria Fortak-Michalska, Bozena_ Szymanska, Wojciech Orlowski, Anna Jurewicz, and Krzysztof Selmaj The molecular mechanisms that regulate the immune function of bone marrow–derived mesenchymal stem cells (BMSCs) are not known.
    [Show full text]
  • Prostacyclin Synthesis by COX-2 Endothelial Cells
    Roles of Cyclooxygenase (COX)-1 and COX-2 in Prostanoid Production by Human Endothelial Cells: Selective Up-Regulation of Prostacyclin Synthesis by COX-2 This information is current as of October 2, 2021. Gillian E. Caughey, Leslie G. Cleland, Peter S. Penglis, Jennifer R. Gamble and Michael J. James J Immunol 2001; 167:2831-2838; ; doi: 10.4049/jimmunol.167.5.2831 http://www.jimmunol.org/content/167/5/2831 Downloaded from References This article cites 36 articles, 23 of which you can access for free at: http://www.jimmunol.org/content/167/5/2831.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 2, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Roles of Cyclooxygenase (COX)-1 and COX-2 in Prostanoid Production by Human Endothelial Cells: Selective Up-Regulation of Prostacyclin Synthesis by COX-21 Gillian E. Caughey,2* Leslie G.
    [Show full text]
  • Studies of Prostaglandin E Formation in Human Monocytes
    Faculty of Technology and Science Biomedical Sciences Sofia Karlsson Studies of prostaglandin E2 formation in human monocytes Karlstad University Studies 2009:43 Sofia Karlsson Studies of prostaglandin E2 formation in human monocytes Karlstad University Studies 2009:43 Sofia Karlsson Studies of prostaglandin E2 formation in human monocytes Licentiate thesis Karlstad University Studies 2009:43 ISSN 1403-8099 ISBN 978-91-7063-266-2 © The Author Distribution: Faculty of Technology and Science Biomedical Sciences SE-651 88 Karlstad +46 54 700 10 00 www.kau.se Printed at: Universitetstryckeriet, Karlstad 2009 ABSTRACT Prostaglandin (PG) E 2 is an eicosanoid derived from the polyunsaturated twenty carbon fatty acid arachidonic acid (AA). PGE 2 has physiological as well as pathophysiological functions and is known to be a key mediator of inflammatory responses. Formation of PGE 2 is dependent upon the activities of three specific enzymes involved in the AA cascade; phospholipase A 2 (PLA 2), cyclooxygenase (COX) and PGE synthase (PGEs). Although the research within this field has been intense for decades, the regulatory mechanisms concerning the PGE 2 synthesising enzymes are not completely established. PGE 2 was investigated in human monocytes with or without lipopolysaccharide (LPS) pre-treatment followed by stimulation with calcium ionophore, opsonised zymosan or phorbol myristate acetate (PMA). Cytosolic PLA 2α (cPLA 2α) was shown to be pivotal for the mobilization of AA and subsequent formation of PGE 2. Although COX-1 was constitutively expressed, monocytes required expression of COX-2 protein in order to convert the mobilized AA into PGH 2. The conversion of PGH 2 to the final product PGE 2 was to a large extent due to the action of microsomal PGEs-1 (mPGEs-1).
    [Show full text]
  • Prostaglandin Transporter OATP2A1/SLCO2A1 Is Essential for Body Temperature Regulation During Fever
    5584 • The Journal of Neuroscience, June 13, 2018 • 38(24):5584–5595 Neurobiology of Disease Prostaglandin Transporter OATP2A1/SLCO2A1 Is Essential for Body Temperature Regulation during Fever Yoshinobu Nakamura,1 XTakeo Nakanishi,1 Hiroaki Shimada,1,2 Junya Shimizu,1 Rika Aotani,1 Shio Maruyama,1 Kei Higuchi,3 Takashi Okura,3 Yoshiharu Deguchi,3 and Ikumi Tamai1 1Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical, and Health Sciences, Kanazawa University, Kanazawa 920-1192, Japan, 2Faculty of Pharmacy, Kindai University, Osaka 577-8502, Japan, and 3Faculty of Pharma-Sciences, Teikyo University, Tokyo 173-8605, Japan Prostaglandin E2 (PGE2 ) in the hypothalamus is a principal mediator of the febrile response. However, the role of organic anion trans- porting polypeptide 2A1 (OATP2A1/SLCO2A1), a prostaglandin transporter, in facilitating this response is unknown. Here, we investi- gated the effect of Slco2a1 deficiency on the body core temperature (Tc) and on the PGE2 concentration in hypothalamus interstitial fluid ␮ Ϫ/Ϫ (Cisf ) and CSF (Ccsf ) of lipopolysaccharide (LPS; 100 g/kg, i.p.)-treated mice of both sexes. Slco2a1 mice did not develop a febrile ϩ/ϩ Ϫ/Ϫ Ϫ/Ϫ response. Ccsf was increased in Slco2a1 and Slco2a1 mice, and Ccsf of Slco2a1 mice was well maintained at 5 h after LPS ϩ/ϩ injection (1160 pg/ml) compared with Slco2a1 mice (316 pg/ml). A microdialysis study revealed that Cisf peaked at 2 h after LPS ϩ/ϩ Ϫ/Ϫ injection in Slco2a1 mice (841 pg/ml), whereas the increase in Cisf was negligible in Slco2a1 mice. The PGE2 plasma concentration in Slco2a1 Ϫ/Ϫ mice (201 pg/ml) was significantly higher than that in Slco2a1 ϩ/ϩ mice (54 pg/ml) at 1 h after LPS injection, whereas the two groups showed similar PGE2 concentrations in the hypothalamus.
    [Show full text]
  • Role of Microsomal Prostaglandin E Synthase 1 in the Kidney
    Role of Microsomal Prostaglandin E Synthase 1 in the Kidney Helene Francois,* Carie Facemire,* Anil Kumar,* Laurent Audoly,† Beverly Koller,‡ and Thomas Coffman* *Divisions of Nephrology, Duke University and Durham Veterans Affairs Medical Centers, Durham, North Carolina, †MedImmune, Gaithersburg, Maryland, and ‡Department of Genetics, University of North Carolina, Chapel Hill, North Carolina Prostaglandin E2 (PGE2) is one of the most ubiquitous prostanoids in the kidney, where it may influence a wide range of physiologic functions. PGE2 is generated through enzymatic metabolism of prostanoid endoperoxides by specific PGE synthases (PGES). Several putative PGES have been identified and cloned, including the membrane-associated, inducible microsomal PGES1 (mPGES1), which is expressed in the kidney. To evaluate the physiologic role of mPGES1 in the kidney, mice with targeted disruption of mPges1 gene were studied, with a focus on responses where PGE2 has been implicated, including urinary concentration, regulation of blood pressure, and response to a loop diuretic. The absence of mPGES1 was associated with a 50% decrease in basal excretion of PGE2 in urine (P < 0.001). In female but not male mPGES1-deficient mice, there was a reciprocal increase in basal excretion of other prostanoids. Nonetheless, urinary osmolalities were similar in mPges1؉/؉ and mPges1؊/؊ mice at baseline and after 12 h of water deprivation. Likewise, there were no differences in blood pressure between mPGES1-deficient and wild-type mice on control or high- or low-salt diets. The furosemide-induced increase in urinary PGE2 excretion that was seen in wild-type mice was attenuated in mPGES1-deficient mice. However, furosemide-associated diuresis was reduced only in male, not female, mPGES1-deficient mice.
    [Show full text]
  • Role of Microsomal Prostaglandin E Synthase-1 (Mpges1)-Derived PGE2 in Patency of the Ductus Arteriosus in the Mouse
    0031-3998/08/6405-0523 Vol. 64, No. 5, 2008 PEDIATRIC RESEARCH Printed in U.S.A. Copyright © 2008 International Pediatric Research Foundation, Inc. Role of Microsomal Prostaglandin E Synthase-1 (mPGES1)-Derived PGE2 in Patency of the Ductus Arteriosus in the Mouse BARBARA BARAGATTI, DARIA SODINI, SATOSHI UEMATSU, AND FLAVIO COCEANI Scuola Superiore Sant’Anna and Institute of Clinical Physiology CNR [B.B., D.S., F.C.], Pisa 56100, Italy; Department of Host Defence [S.U.], Research Institute for Microbial Diseases, Osaka University, Osaka 565-0871, Japan ABSTRACT: Prostaglandin E2 (PGE2) plays a key role in the ductus close or even equal in magnitude to that of a dual COX1/ arteriosus, prenatally by maintaining patency and postnatally by COX2 inhibitor (7–9); the greater impairment of PGE2-based promoting tissue remodeling for closure. Here, by using near-term relaxation upon COX2 than COX1 deletion (5) along with the mouse fetuses with (wild-type, WT) and without microsomal PGE marked curtailment of PGE formation after inhibition of Ϫ Ϫ 2 synthase-1 (mPGES1 / ), we have examined the importance of this either COX2 or mPGES1 (6,7). Consistent with the preferen- enzyme for PGE formation and function. mPGES1Ϫ/Ϫ ductus, 2 tial operation of the COX2/mPGES1 route for PGE synthesis unlike WT ductus, contracted little, or not all, to indomethacin in 2 G is also evidence from adult blood vessels (10) and the notion that vitro. Coincidentally, as evident from responses to N -nitro-L- arginine methyl ester and zinc photoporphyrin, the mutant showed no mPGES1 is catalytically most active among the PGES (11).
    [Show full text]