DOCSLIB.ORG

Resolution of Prostaglandin Endoperoxide Synthase

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Resolution of Prostaglandin Endoperoxide Synthase Proc. Nati. Acad. Sci. USA Vol. 74, No. 9, pp. 3691-3695, September 1977 Biochemistry Resolution of prostaglandin endoperoxide synthase and thromboxane synthase of human platelets (subcellular distribution/12L-hydroperoxy-5,8,10,14-eicosatetraenoic acid/Triton X-100/DEAE-cellulose) SVEN HAMMARSTROM AND PIERRE FALARDEAU* Department of Chemistry, Karolinska Institutet, S-104 01 Stockholm, Sweden Communicated by Hugo Theorell, June 13, 1977 ABSTRACT Thromboxane synthase was localized to the icated human platelets (compare below) sedimenting between microsomes of human platelets. The enzyme was insensitive to 100,000 X g (1 hr) and 300,000 X g (4 hr), 20 mg of protein, sulfhydryl reagents and thiols but was inhibited by 12L-hy- droperoxy-5,8,10,14-eicosatetraenoic acid (concentration for were incubated at 370 for 8 min in 15 ml of 50 mM Tris-HCI, 50% inhibition = 0.1 mM). Treatment of microsomes with Tri- pH 7.4/1 mM 2,2'-dipyridyl. The product eluting from silicic ton X-100 solubilized the enzymes that catalyze the conversion acid with diethyl ether/light petroleum (15:85, vol/vol) was of arachidonic acid to thromboxane B2. The solubilized material homogeneous as the methyl ester on thin-layer chromatography was resolved by DEAE-cellulose chromatography into two [RF = 0.64; solvent: diethyl ether/light petroleum (35:65, vol/ components, one converting arachidonic acid to prostaglandins vol)]. SnC12 reduction converted this product to 12L-hy- G2 and H2 and the other converting prostaglandin H2 to droxy-5,8,10,14-eicosatetraenoic acid (RF = 0.41) as judged by thromboxane B2. gas-liquid chromatography/mass spectrometry (4). Thromboxanes are a novel class of compounds derived from Washed Human Platelets and Subcellular Fractionation. prostaglandin endoperoxides (1). They are extremely potent Blood was collected in 77 mM EDTA, 7.5%, vol/vol, from do- as inducers of the platelet release reaction and aggregation (1) nors who had not taken drugs for at least 1 week. Washed and as stimulators of smooth muscle contractions (2, 3). platelets were prepared as described (12). They were suspended Thromboxane A2 is the major component of rabbit aorta con- in 50 mM Tris.HCI, pH 7.4, or in 10 mM potassium phosphate, tracting substance [RCS (2)]. It contains an unstable oxane ox- pH 7.4 (10 and 5 ml/400 ml of blood, respectively), disrupted etane structure and reacts rapidly with water (t1/2 at 370 = 32 by sonication with a Branson model S-125 sonifier (six 5-sec sec) to form a stable derivative (1). This derivative, 8-(1-hy- treatments with 1-min intervals for cooling; power setting: 4) droxy-3-oxopropyl)-9,12L-dihydroxy-5,10-heptadecadienoic or by hypotonic lysis after glycerol loading (13), fractionated acid (thromboxane B2), was originally discovered as a product by differential centrifugation (1,900 X g, 15 min; 12,000 X g, of arachidonic acid metabolism in human platelets (4). An en- 15 min; and 100,000 X g, 1 hr). Plasma membranes (14), dense zyme catalyzing the conversion of prostaglandin endoperoxides bodies, and a granules (15) were isolated as described. Platelet to thromboxanes has been described in platelet microsomes (1, disruption and subsequent manipulations were performed at 5-11). This paper deals with the solubilization of the enzymes 0-4g. in human platelets that convert arachidonic acid to throm- Characterization of Products Formed from PGG2 and boxane A2 and their resolution into prostaglandin endoperoxide PGH2. A mixture of 625 nmol of [1-14C]PGG2 or [1-14C]PGH2 synthase and thromboxane synthase components. (0.6 Ci/mol), 1 mmol of Tris-HCI, pH 7.4, and 30 mg of mi- crosomal protein (12,000 X g-100,000 X g sediment) in a total volume of 20 ml was incubated at 370 for 20 sec. The reaction MATERIALS AND METHODS was stopped by adding 100 ml of SnC12 (5 mg/ml) in dioxane [1-14C]Arachidonic acid (50-60 Ci/mol, The Radiochemical to reduce untransformed endoperoxide to PGF2 (12). Products Centre, Amersham, England) and arachidonic acid (Nu Chek were extracted at pH 3 with diethyl ether, methylated, and Prep., Inc., Elysian, MN) were used for enzyme assays and to separated by preparative thin-layer chromatography (solvent: prepare [1-14C]prostaglandin (PG) G2 and [1-14C]PGH2 (1 diethyl ether/methanol, 49:1, vol/vol). After elution from the Ci/mol) (12). Nordihydroguaiaretic acid was from Bast's Suc- plate with ethyl acetate/methanol (1:1) and conversion to cessors, Ltd., Copenhagen. Thromboxane B2 and prostaglandins methoxime (PGE2), trimethylsilyl derivatives, the following were kindly supplied by Upjohn, Kalamazoo, MI; L-cysteine, compounds were identified by gas-liquid chromatography/ 2,2'-dipyridyl, dithiothreitol, ethylenediamine tetraacetic acid mass spectrometry: 12L-hydroxy-5,8,10-heptadecatrienoic acid (EDTA), ethanethiol, N-ethylmaleimide, beef blood hemo- (HHT), thromboxane B2, PGE2, and PGF2a. globin (Type I), p-hydroxymercuribenzoic acid, 2-mercapto- Enzyme Assays. Incubations with arachidonic acid were ethanol, SnCl2, and L-tryptophan were from Sigma; DEAE- performed in one of two ways: (a) A reaction mixture (0.5 ml) cellulose (DE-52) was from Whatman; silica gel G and Tris were containing [1-14C]arachidonic acid (0.7 Ci/mol; 33 or 66 nmol), from Merck; and silicic acid was from Mallinckrodt. 12L- Tris-HCI at pH 7.4 (25 ,mol), and a platelet subcellular fraction Hydroxy-5,8,10,14-eicosatetraenoic acid (HETE) was prepared was incubated at 37° for 45 sec. Ethanol (2.5 ml) and water (2.5 as described (4). ml) were added, and the mixture was acidified to pH 3 and 12L-Hydroperoxy-5,8,10,14-eicosatetraenoic Acid extracted with diethyl ether. Thin-layer chromatography of (HPETE). Arachidonic acid (2.5 mg) and a fraction from son- Abbreviations: HETE, 12L-hydroxy-5,810,14-eicosatetraenoic acid; HPETE, 12L-hydroperoxy-5,8,10,14-eicosatetraenoic acid; HHT, The costs of publication of this article were defrayed in part by the 12L-hydroxy-5,8,10-heptadecatrienoic acid; PG, prostaglandin; TX, payment of page charges. This article must therefore be hereby marked thromboxane. "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate * Present address: Department of Nephrology, Centre Hospitalier this fact. Universitaire, Sherbrooke, PQ, Canada, J1H 5N4. 3691 Downloaded by guest on September 30, 2021 3692 Biochemistry: Hammarstrom and Falardeau Proc. Natl. Acad. Sci. USA 74 (1977) 4 I 100- I I I 80- I I I 02-- I I a 201 I I 60- % I I + I I I LALC~I40- 0 I .cc0 a- I NX, 8R 60( I -x 20- -X- I I I I I 30 +86 nmot / 15 10 5 0 PGG2,( Distance from origin (cm) Z 20- 2 )( FIG. 1. Thin-layer radiochromatograms of products formed from E // [1-14CJPGG2 during incubations (370, 20 sec) with native (Upper) c (' or boiled (Lower) platelet microsomes. Excess PGG2 was reduced to PGF20 with SnCl2 at the end of the incubation. 12-OH-17:3, 12L- I hydroxy-5,8,10-heptadecatrienoic acid (HHT); TXB2, thromboxane m 1, B2. Solvent system: diethyl ether/methanol, 49:1, vol/vol. xi-10- +43mPt ++3 nmol PGO'2 methyl esters was performed with diethyl ether/light petro- leum, 3:7 (vol/vol) as solvent. (b) A reaction mixture (0.1 ml) containing [1-'4C]arachidonic acid (55 Ci/mol; 2 nmol), he- moglobin (0.2 nmol), L-tryptophan (0.5 ,mol), Tris-HCI at pH 7.4 (10 ,umol), and microsomes, solubilized proteins, or aliquots 5 1015 30 of eluates from DEAE-cellulose chromatography was incubated Seconds for 5 min at 240. Citric acid (0.1 M in water, 1 ml) and throm- FIG. 2. Time course of PGG2 disappearance (Upper) and boxane B2 (10-gg) were added and the mixture was extracted thromboxane B2 formation (Lower). [1-14C]PGG2, 72.5 AM (0 O) with diethyl ether. After washing and methylation, the ether or 145 AM (X- - -X), was incubated with platelet microsomes at 37°. Incubations were stopped with SnCl2 in dioxane and products were extracts were analyzed by thin-layer chromatography in two separated by thin-layer chromatography as shown in Fig. 1. A second solvent systems: ethyl acetate/2,2,4-trimethylpentane/water, addition of substrate was made after 15 sec (arrows). 5:10:10, vol/vol/vol (I) and diethyl ether/methanol, 49:1, vol/vol (II). Assays with were mixture (0.6 ml) contained [1-'4C]PGG2 (0.6 Ci/moi; 43 or 86 PGG2 performed as follows: the reaction nmol), Tris.HCI at pH 7.4 (30,umol), and a platelet subcellular fraction. After 15 or 20 sec at 370, the reaction was stopped with Table 1. Effects of sulfhydryl reagents, thiols, chelators, dioxane (3 ml) containing SnCl2 (5 mg/ml). After extraction nordihydroguaiaretic acid, HPETE, and HETE on thromboxane and methylation, products were analyzed by thin-layer chro- B2 formation by platelet microsomes matography with solvent II. Microsomes, solubilized micro- Concentration, % inhibition of somes, and eluates from DEAE-cellulose columns were assayed Substance mM TXB2 formation* with PGH2 as substrate. The reaction mixture (0.1 ml) con- taining [1-14CJPGH2 (12 nmol; 1 Ci/mol) Tris-HCl at pH 7.4 Nordihydroguaiaretic 0.1 0 (10,mol), and enzyme was incubated at 240 for 1 min. The acid 0.3 50 reaction was terminated as described above for incubations with 0.5 80 arachidonic HPETE 0.1 50 acid (procedure b). Products were analyzed by 0.2 80 thin-layer chromatography with solvent II. HETE 0.3 15 Thin-layer chromatograms were scanned for radioactivity p-Hydroxymercuri- 1 20 with a Berthold Dunnschicht Scanner II. Thromboxane B2 benzoic acid added as carrier was detected by spraying plates with phos- N-Ethylmaleimide 1 0 phomolybdic acid and heating them briefly.
Load more

Recommended publications
  • Simultaneous Determination of PUFA-Derived Pro-Resolving Metabolites and Pathway Markers Using Chiral Chromatography and Tandem
    Simultaneous determination of PUFA-derived pro-resolving metabolites and pathway markers using chiral chromatography and tandem mass spectrometry Andy Toewe, Laurence Balas, Thierry Durand, Gerd Geisslinger, Nerea Ferreirós To cite this version: Andy Toewe, Laurence Balas, Thierry Durand, Gerd Geisslinger, Nerea Ferreirós. Simultaneous determination of PUFA-derived pro-resolving metabolites and pathway markers using chiral chro- matography and tandem mass spectrometry. Analytica Chimica Acta, Elsevier Masson, 2018, 1031, pp.185-194. 10.1016/j.aca.2018.05.020. hal-02405326 HAL Id: hal-02405326 https://hal.archives-ouvertes.fr/hal-02405326 Submitted on 11 Dec 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Simultaneous determination of PUFA-derived pro-resolving metabolites and pathway markers using chiral chromatography and tandem mass spectrometry a b b a, c a, * Andy Toewe , Laurence Balas , Thierry Durand , Gerd Geisslinger , Nerea Ferreiros a pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe-University Frankfurt, Germany b Institut des Biomolecules Max Mousseron, IBMM, UMR 5247, University of Montpellier, CNRS, ENSCM, Montepellier, France c Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group TMP, Frankfurt, Germany abstract Lipid mediators play an important role as biological messengers involved in inflammatory processes.
    [Show full text]
  • Serum Malondialdehyde Is Associated with Non-Alcoholic Fatty Liver and Related Liver Damage Differentially in Men and Women
    antioxidants Article Serum Malondialdehyde is Associated with Non-Alcoholic Fatty Liver and Related Liver Damage Differentially in Men and Women Shira Zelber-Sagi 1,2,*, Dana Ivancovsky-Wajcman 1, Naomi Fliss-Isakov 2,3, Michal Hahn 4, 2,3 2,3 2,3, 4, Muriel Webb , Oren Shibolet , Revital Kariv y and Oren Tirosh y 1 School of Public Health, University of Haifa, Haifa 3498838, Israel; [email protected] 2 Department of Gastroenterology, Tel Aviv Medical Center, Tel Aviv 6423914, Israel; naomifl@tlvmc.gov.il (N.F.-I.); [email protected] (M.W.); [email protected] (O.S.); [email protected] (R.K.) 3 Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel 4 Institute of Biochemistry, Food Science and Nutrition, The RH Smit Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rechovot 76100001, Israel; [email protected] (M.H.); [email protected] (O.T.) * Correspondence: [email protected]; Tel.: +972-3-6973984 The last two authors equally contributed to the paper. y Received: 25 May 2020; Accepted: 25 June 2020; Published: 2 July 2020 Abstract: Background: Non-alcoholic fatty liver disease (NAFLD) and steatohepatitis (NASH) are associated with increased oxidative stress and lipid peroxidation, but large studies are lacking. The aim was to test the association of malondialdehyde (MDA), as a marker of oxidative damage of lipids, with NAFLD and liver damage markers, and to test the association between dietary vitamins E and C intake and MDA levels. Methods: A cross-sectional study was carried out among subjects who underwent blood tests including FibroMax for non-invasive assessment of NASH and fibrosis.
    [Show full text]
  • Diclofenac Enhances Docosahexaenoic Acid-Induced Apoptosis in Vitro in Lung Cancer Cells
    cancers Article Diclofenac Enhances Docosahexaenoic Acid-Induced Apoptosis in Vitro in Lung Cancer Cells Rosemary A. Poku 1, Kylee J. Jones 2, Megan Van Baren 2 , Jamie K. Alan 3 and Felix Amissah 2,* 1 Department of Foundational Sciences, College of Medicine, Central Michigan University, Warriner Hall, 319, Mt Pleasant, MI 48859, USA; [email protected] 2 Department of Pharmaceutical Sciences, Ferris State University, College of Pharmacy, 220 Ferris Dr, Big Rapids, MI 49307, USA; [email protected] (K.J.J.); [email protected] (M.V.B.) 3 Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI 48824, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-231-591-3790 Received: 9 September 2020; Accepted: 17 September 2020; Published: 20 September 2020 Simple Summary: Polyunsaturated fatty acids (PUFAs) and non-steroidal anti-inflammatory drugs (NSAIDs) have limited anticancer capacities when used alone. We examined whether combining NSAIDs with docosahexaenoic (DHA) would increase their anticancer activity on lung cancer cell lines. Our results indicate that combining DHA and NSAIDs increased their anticancer activities by altering the expression of critical proteins in the RAS/MEK/ERK and PI3K/Akt pathways. The data suggest that DHA combined with low dose diclofenac provides more significant anticancer potential, which can be further developed for chemoprevention and adjunct therapy in lung cancer. Abstract: Polyunsaturated fatty acids (PUFAs) and non-steroidal anti-inflammatory drugs (NSAIDs) show anticancer activities through diverse molecular mechanisms. However, the anticancer capacities of either PUFAs or NSAIDs alone is limited. We examined whether combining NSAIDs with docosahexaenoic (DHA), commonly derived from fish oils, would possibly synergize their anticancer activity.
    [Show full text]
  • Antioxidants and Second Messengers of Free Radicals
    antioxidants Antioxidants and Second Messengers of Free Radicals Edited by Neven Zarkovic Printed Edition of the Special Issue Published in Antioxidants www.mdpi.com/journal/antioxidants Antioxidants and Second Messengers of Free Radicals Antioxidants and Second Messengers of Free Radicals Special Issue Editor Neven Zarkovic MDPI • Basel • Beijing • Wuhan • Barcelona • Belgrade Special Issue Editor Neven Zarkovic Rudjer Boskovic Institute Croatia Editorial Office MDPI St. Alban-Anlage 66 4052 Basel, Switzerland This is a reprint of articles from the Special Issue published online in the open access journal Antioxidants (ISSN 2076-3921) from 2018 (available at: https://www.mdpi.com/journal/ antioxidants/special issues/second messengers free radicals) For citation purposes, cite each article independently as indicated on the article page online and as indicated below: LastName, A.A.; LastName, B.B.; LastName, C.C. Article Title. Journal Name Year, Article Number, Page Range. ISBN 978-3-03897-533-5 (Pbk) ISBN 978-3-03897-534-2 (PDF) c 2019 by the authors. Articles in this book are Open Access and distributed under the Creative Commons Attribution (CC BY) license, which allows users to download, copy and build upon published articles, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. The book as a whole is distributed by MDPI under the terms and conditions of the Creative Commons license CC BY-NC-ND. Contents About the Special Issue Editor ...................................... vii Preface to ”Antioxidants and Second Messengers of Free Radicals” ................ ix Neven Zarkovic Antioxidants and Second Messengers of Free Radicals Reprinted from: Antioxidants 2018, 7, 158, doi:10.3390/antiox7110158 ...............
    [Show full text]
  • Role of Thrombin and Thromboxane A2 in Reocclusion Following Coronary
    Proc. Natl. Acad. Sci. USA Vol. 86, pp. 7585-7589, October 1989 Medical Sciences Role of thrombin and thromboxane A2 in reocclusion following coronary thrombolysis with tissue-type plasminogen activator (thrombolytic therapy/coronary thrombosis/platelet activation/reperfusion) DESMOND J. FITZGERALD*I* AND GARRET A. FITZGERALD* Divisions of *Clinical Pharmacology and tCardiology, Vanderbilt University, Nashville, TN 37232 Communicated by Philip Needleman, June 28, 1989 (receivedfor review April 12, 1989) ABSTRACT Reocclusion of the coronary artery occurs against the prothrombinase formed on the platelet surface after thrombolytic therapy of acute myocardlal infarction (13) and the neutralization ofheparin by platelet factor 4 (14) despite routine use of the anticoagulant heparin. However, and thrombospondin (15), proteins released by activated heparin is inhibited by platelet activation, which is greatly platelets. enhanced in this setting. Consequently, it is unclear whether To address the role of thrombin during coronary throm- thrombin induces acute reocclusion. To address this possibility, bolysis, we have examined the effect of a specific thrombin we examined the effect of argatroban {MCI9038, (2R,4R)- inhibitor, argatroban {MCI9038, (2R,4R)-4-methyl-1-[N-(3- 4-methyl-l-[Na-(3-methyl-1,2,3,4-tetrahydro-8-quinolinesulfo- methyl-1,2,3,4-tetrahydro-8-quinolinesulfonyl)-L-arginyl]-2- nyl)-L-arginyl]-2-piperidinecarboxylic acid}, a specific throm- piperidinecarboxylic acid} on the response to tissue plasmin- bin inhibitor, on the response to tissue-type plasminogen ogen activator (t-PA) in a closed-chest canine model of activator in a dosed-chest canine model of coronary thrombo- coronary thrombosis. MCI9038, an argimine derivative which sis. MCI9038 prolonged the thrombin time and shortened the binds to a hydrophobic pocket close to the active site of time to reperfusion (28 + 2 min vs.
    [Show full text]
  • AA Metabolism, 139 A2b1, 69, 70, 318–323 ABCB1 Gene, 176 Abciximab
    Index A aIIbb3, 60, 62, 69, 70, 73, 74, 79, 80 AA. See Arachidonic acid (AA) AJW202, 293 AA metabolism, 139 Akt-1, 345 a2b1, 69, 70, 318–323 Akt-2, 345 ABCB1 gene, 176 ALX-0081, 296 Abciximab, 206–208, 497, 500, 501, 504–505, ALX-0681, 298 509, 513, 514, 525 Anagrelide, 227, 229 approval, 206 Ankle brachial indexes (ABIs), 549, 587, EPIC trial, 207 595–597 EPILOG study, 207 framingham risk score, 551 EPISTENT study, 207 screening, 550 platelet binding, 207 sensitivity, 551 thrombocytopenia, 206 specificity, 551 in unstable angina, 208 vascular risk, 550 ABIs. See Ankle brachial indexes (ABIs) Antagomirs, 439 Acetylsalicylic acid (aspirin), 137–158 Antibodies, 317 ACS. See Acute coronary syndromes (ACS) Anticoagulants, 525, 537 Acute coronary syndromes (ACS), 288 Anticoagulation, 533, 534 Acute ischemic stroke, 143, 149–150 Antioxidant, 233 Acute myocardial infarction, 142, 143, 149, Antiplatelet agents, 263, 553 153, 157 Antithrombotic Trialist’s ADAMTS-13, 93 Collaboration, 553 Adenosine diphosphate (ADP), 37, 88, 96, aspirin, 553 166, 473 clopidogrel, 553 Adenosine triphosphate (ATP), 37 dipyridamole, 553 Adhesion, 90, 112–118, 125 meta-analysis, 560 Adhesion molecules infiltration, 264 picotamide, 553 Adiponectin, 315 risk reduction, 553 ADP. See Adenosine diphosphate (ADP) ticlopidine, 553 ADP inhibitors (or P2Y12 blockers), 497–499, vascular events, 553 501–505, 507–509, 513, 514 Antiplatelet therapy, 472 ADP-receptor antagonists, 169 Apixaban, 535, 539 Adverse effects, 153–156 Aptamers, 299–301 Aegyptin, 327 Arachidonic acid (AA), 474 P. Gresele et al. (eds.), Antiplatelet Agents, Handbook of Experimental 607 Pharmacology 210, DOI 10.1007/978-3-642-29423-5, # Springer-Verlag Berlin Heidelberg 2012 608 Index ARC1779, 299–300 TXS signaling, 279 ARC15105, 300 Cardiovascular death, 248 AR-C69931MX, 456 Carotid endarterectomy (ACE) inhibition, Aspirin, 474, 496, 498–499, 501–502, 143, 156 504–507, 511–514, 520, 522, Carotid stenosis, 523 527, 531, 533–537, 539, CCBs.
    [Show full text]
  • OBE022, an Oral and Selective Prostaglandin F2α Receptor Antagonist As an Effective and Safe Modality for the Treatment of Pret
    Supplemental material to this article can be found at: http://jpet.aspetjournals.org/content/suppl/2018/05/18/jpet.118.247668.DC1 1521-0103/366/2/349–364$35.00 https://doi.org/10.1124/jpet.118.247668 THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS J Pharmacol Exp Ther 366:349–364, August 2018 Copyright ª 2018 by The American Society for Pharmacology and Experimental Therapeutics OBE022, an Oral and Selective Prostaglandin F2a Receptor Antagonist as an Effective and Safe Modality for the Treatment of Preterm Labor s Oliver Pohl, André Chollet, Sung Hye Kim, Lucia Riaposova, François Spézia, Frédéric Gervais, Philippe Guillaume, Philippe Lluel, Murielle Méen, Frédérique Lemaux, Vasso Terzidou, Phillip R. Bennett, and Jean-Pierre Gotteland ObsEva SA, Plan-les-Ouates, Geneva, Switzerland (O.P., A.C., J.-P.G.); Imperial College London, Parturition Research Group, Institute of Reproductive and Developmental Biology,HammersmithHospitalCampus,EastActon,London,UnitedKingdom(S.H.K.,L.R., V.T., P.R.B.); Citoxlab, Evreux, France (F.S., F.G.); Porsolt Research Laboratory, Le Genest-Saint-Isle, France (P.G.); Urosphere SAS, Toulouse, France (P.L., M.M.); BioTrial, Rennes, France (F.L.); and André Chollet Consulting, Tannay, Switzerland (A.C.) Downloaded from Received February 26, 2018; accepted May 15, 2018 ABSTRACT Preterm birth is the major challenge in obstetrics, affecting aggregation. In in vitro studies, OBE002 inhibited sponta- ∼ 10% of pregnancies. Pan-prostaglandin synthesis inhibitors neous, oxytocin- and PGF2a-induced human myometrial jpet.aspetjournals.org [nonsteroidal anti-inflammatory drugs (NSAIDs)] prevent preterm contractions alone and was more effective in combination labor and prolong pregnancy but raise concerns about fetal renal with atosiban or nifedipine.
    [Show full text]
  • Oxisresearch™ a Division of OXIS Health Products, Inc
    OxisResearch™ A Division of OXIS Health Products, Inc. BIOXYTECHÒ MDA-586™ Spectrophotometric Assay for Malondialdehyde For Research Use Only. Not For Use In Diagnostic Procedures. Catalog Number 21044 INTRODUCTION The Analyte Lipid peroxidation is a well-established mechanism of cellular injury in both plants and animals, and is used as an indicator of oxidative stress in cells and tissues. Lipid peroxides, derived from polyunsaturated fatty acids, are unstable and decompose to form a complex series of compounds. These include reactive carbonyl compounds, of which the most abundant is malondialdehyde (MDA). Therefore, measurement of malondialdehyde is widely used as an indicator of lipid peroxidation (1). Increased levels of lipid peroxidation products have been associated with a variety of chronic diseases in both humans (2, 3) and model systems (4, 5). MDA reacts readily with amino groups on proteins and other biomolecules to form a variety of adducts (1), including cross-linked products (6). MDA also forms adducts with DNA bases that are mutagenic (7, 8) and possibly carcinogenic (9). DNA-protein cross-links are another result of the reaction between DNA and MDA (10). The TBARS method is commonly used to measure MDA in biological samples (11). However, this reaction is relatively nonspecific; both free and protein-bound MDA can react. The MDA-586 method is designed to assay free MDA or, after a hydrolysis step, total MDA (i.e., free and protein-bound Schiff base conjugates). The assay conditions serve to minimize interference from other lipid peroxidation products, such as 4- hydroxyalkenals. PRINCIPLES OF THE PROCEDURE The MDA-586 method1 (12) is based on the reaction of a chromogenic reagent, N-methyl-2- phenylindole (R1, NMPI), with MDA at 45°C.
    [Show full text]
  • Neuroprotective Effects of Geniposide from Alzheimer's Disease Pathology
    Neuroprotective effects of geniposide from Alzheimer’s disease pathology WeiZhen Liu1, Guanglai Li2, Christian Hölscher2,3, Lin Li1 1. Key Laboratory of Cellular Physiology, Shanxi Medical University, Taiyuan, PR China 2. Second hospital, Shanxi medical University, Taiyuan, PR China 3. Neuroscience research group, Faculty of Health and Medicine, Lancaster University, Lancaster LA1 4YQ, UK running title: Neuroprotective effects of geniposide corresponding author: Prof. Lin Li Key Laboratory of Cellular Physiology, Shanxi Medical University, Taiyuan, PR China Email: [email protected] Neuroprotective effects of geniposide Abstract A growing body of evidence have linked two of the most common aged-related diseases, type 2 diabetes mellitus (T2DM) and Alzheimer disease (AD). It has led to the notion that drugs developed for the treatment of T2DM may be beneficial in modifying the pathophysiology of AD. As a receptor agonist of glucagon- like peptide (GLP-1R) which is a newer drug class to treat T2DM, Geniposide shows clear effects in inhibiting pathological processes underlying AD, such as and promoting neurite outgrowth. In the present article, we review possible molecular mechanisms of geniposide to protect the brain from pathologic damages underlying AD: reducing amyloid plaques, inhibiting tau phosphorylation, preventing memory impairment and loss of synapses, reducing oxidative stress and the chronic inflammatory response, and promoting neurite outgrowth via the GLP-1R signaling pathway. In summary, the Chinese herb geniposide shows great promise as a novel treatment for AD. Key words: Alzheimer’s disease, geniposide, amyloid-β, neurofibrillary tangles, oxidative stress, inflammatation, type 2 diabetes mellitus, glucagon like peptide receptor, neuroprotection, tau protein Neuroprotective effects of geniposide 1.
    [Show full text]
  • Eicosanoids & Platelet Activating Factor
    Eicosanoids & Platelet Activating Factor Prof. Sanjay Khattri Dept. of Pharmacology & Therapeutics King George’s Medical University, Lucknow 1 Autacoid These are the substances produced by wide variety of cells that act locally at the site of production. (local hormones) 2 Mediators of Inflammation and Immune reaction 1. Vasoactive amines (Histamine and Serotonin) 2.Eicosanoids 3.Platlet Activating Factor 4.Bradykinins 4.Nitric Oxide 5.Neuropeptides 6.Cytokinines 3 EICOSANOIDS PGs, TXs and LTs are all derived from eicosa (referring to 20 C atoms) tri/tetra/ penta enoic acids. Therefore, they can be collectively called eicosanoids. Major source: 5,8,11,14 eicosa tetraenoic acid (arachidonic acid). Other eicosanoids of increasing interest are: lipoxins and resolvins. The term prostanoid encompasses both prostaglandins and thromboxanes. 4 EICOSANOIDS Contd…. In most instances, the initial and rate-limiting step in eicosanoid synthesis is the liberation of intracellular arachidonate, usually in a one-step process catalyzed by the enzyme phospholipase A2 (PLA2). PLA2 generates not only arachidonic acid but also lysoglyceryl - phosphorylcholine (lyso-PAF), the precursor of platelet activating factor (PAF). 5 EICOSANOIDS Contd…. Corticosteroids inhibit the enzyme PLA2 by inducing the production of lipocortins (annexins). The free arachidonic acid is metabolised separately (or sometimes jointly) by several pathways, including the following: Cyclo-oxygenase (COX)- Two main isoforms exist, COX-1 and COX-2 Lipoxygenases- Several subtypes, which
    [Show full text]
  • Role of Arachidonic Acid and Its Metabolites in the Biological and Clinical Manifestations of Idiopathic Nephrotic Syndrome
    International Journal of Molecular Sciences Review Role of Arachidonic Acid and Its Metabolites in the Biological and Clinical Manifestations of Idiopathic Nephrotic Syndrome Stefano Turolo 1,* , Alberto Edefonti 1 , Alessandra Mazzocchi 2, Marie Louise Syren 2, William Morello 1, Carlo Agostoni 2,3 and Giovanni Montini 1,2 1 Fondazione IRCCS Ca’ Granda-Ospedale Maggiore Policlinico, Pediatric Nephrology, Dialysis and Transplant Unit, Via della Commenda 9, 20122 Milan, Italy; [email protected] (A.E.); [email protected] (W.M.); [email protected] (G.M.) 2 Department of Clinical Sciences and Community Health, University of Milan, 20122 Milan, Italy; [email protected] (A.M.); [email protected] (M.L.S.); [email protected] (C.A.) 3 Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Pediatric Intermediate Care Unit, 20122 Milan, Italy * Correspondence: [email protected] Abstract: Studies concerning the role of arachidonic acid (AA) and its metabolites in kidney disease are scarce, and this applies in particular to idiopathic nephrotic syndrome (INS). INS is one of the most frequent glomerular diseases in childhood; it is characterized by T-lymphocyte dysfunction, alterations of pro- and anti-coagulant factor levels, and increased platelet count and aggregation, leading to thrombophilia. AA and its metabolites are involved in several biological processes. Herein, Citation: Turolo, S.; Edefonti, A.; we describe the main fields where they may play a significant role, particularly as it pertains to their Mazzocchi, A.; Syren, M.L.; effects on the kidney and the mechanisms underlying INS. AA and its metabolites influence cell Morello, W.; Agostoni, C.; Montini, G.
    [Show full text]
  • Success of Prostaglandin E2 in Structure–Function Is a Challenge for Structure-Based Therapeutics
    COMMENTARY Success of prostaglandin E2 in structure–function is a challenge for structure-based therapeutics Charles N. Serhan*† and Bruce Levy*‡ *Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesiology, Perioperative and Pain Medicine and ‡Critical Care and Pulmonary Medicine, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115 rostaglandin (PG) E2 is almost ubiquitous in humans and evokes potent diverse actions. Utility is the price of its perfec- Ption. PGE is a founding member of the 2 PGs, a class of mediators that belongs to the still growing family of bioactive au- tacoids known as the eicosanoids (1–3). The main classes include enzymatically generated products such as thrombox- anes, leukotrienes, lipoxins, and EETs, as well as others that are produced via nonenzymatic mechanisms, e.g., isopros- tanes and cyclopentaeone PGs that are increasing in number and appreciation (4, 5). PGE2 regulates key responses in the major human systems including re- productive, gastrointestinal, neuroendo- crine, and immune (Fig. 1). Formed by conversion of arachidonic acid via cyclo- oxygenase (COX) and specific synthases, Fig. 1. Diverse actions of PGE2 and selective targeted biosynthesis in inflammation. (Inset) Eicosanoid family major enzymatic classes of cyclooxygenases and lipoxygenase pathways (see text for details). PGE2 stereospecifically exerts potent (nano- to micromolar range) tissue- and cell type-selective actions (1–6). The importance of PGs in inflammation was from protecting gastrointestinal mucosa electrophoresis that this lipid-soluble brought into view by the discovery of J. to regulating smooth muscle and fever, activity behaved as an acid. Vane and colleagues (7) that nonsteroi- set a steep challenge for designer drug Bergstro¨m’s main research was in bile dal antiinflammatory drugs (NSAIDs) hunters to achieve, namely selectivity acids and steroids.
    [Show full text]