DOCSLIB.ORG

Stimulation by Ceramide of Phospholipase A2 Activation Through a Mechanism Related to the Phospholipase C-Initiated Signaling Pathway in Rabbit Platelets'

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Stimulation by Ceramide of Phospholipase A2 Activation Through a Mechanism Related to the Phospholipase C-Initiated Signaling Pathway in Rabbit Platelets' J. Biochem. 125, 96-102 (1999) Stimulation by Ceramide of Phospholipase A2 Activation through a Mechanism Related to the Phospholipase C-Initiated Signaling Pathway in Rabbit Platelets' Takashi Sato,2 Tadashi Kageura, Tsutomu Hashizume, Misako Hayama, Kazuyuki Kitatani, and Satoshi Akiba Departmentof PathologicalBiochemistry, Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Kyoto, Kyoto 607-8414 ReceivedAugust 7, 1998;accepted September 30, 1998 To study the involvement of sphingolipids in glycerophospholipid metabolism, the contribution of ceramide to the activation of group IV cytosolic phospholipase A2 (cPLA2) was investigated in platelets using cell-permeable C6-ceramide (N-hexanoylsphingosine). The addition of ceramide led to potentiation of thrombin-induced activation of cPLA2 and mitogen-activated protein kinase (MAPK) as well as arachidonic acid release and lysophos phatidyicholine formation. However, ceramide by itself did not induce any response. The arachidonic acid release due to the synergistic action of ceramide and thrombin was inhibited by PD98059, a MAPK kinase inhibitor. Ceramide also stimulated thrombin-in duced protein kinase C (PKC) activation, but ceramide by itself failed to do so. Further more, ceramide synergistically enhanced diacylglycerol (DAG) formation and Ca" mobili zation with thrombin, and also DAG formation with Ca+-ionophore A23187. The DAG formation in response to ceramide with thrombin or A23187, as well as arachidonic acid release with thrombin were completely inhibited by U73122, a phospholipase C (PLC) inhibitor. These results suggest that ceramide triggers PLC activation through its synergis tic action with thrombin, and subsequently potentiates the sequential PKC-MAPK cascade- cPLA2 pathway, thus resulting in enhancement of arachidonic acid release. Key words: ceramide, cytosolic phospholipase A2, mitogen-activated protein kinase, phospholipase C, protein kinase C. Receptor-mediated stimulation of the sphingomyelin path- activation of sphingomyelinase-initiated sphingomyelin way via activation of sphingomyelinase generates biologi metabolism, thus suggesting the occurrence of cross-talk cally active sphingolipid metabolites, such as ceramide and between the glycerophospholipid and sphingomyelin sig sphingosine, which have been shown to act as cellular naling pathways. Indeed, cell-permeable C6-ceramide (N- second messengers in a variety of signal transduction hexanoylsphingosine), sphingosine or sphingomyelinase pathways (reviewed in Refs. 1 and 2). Ceramide, a product treatment has been shown to stimulate PLD activation in of sphingomyelin hydrolysis, is known to be generated Swiss 3T3 fibroblasts (6), neural-derived NG 108-15 cells following stimulation with cytokines, growth factors, FAS (7), and human fibroblasts (8), whereas C2-(N-acetyl ligand, and vitamin D3, and is widely accepted to exhibit sphingosine) or C6-ceramides prevent the activation in rat many biological activities, including cell proliferation, or human fibroblasts (9-12), rat basophilic leukemia cells differentiation and apoptosis, in a variety of cells (reviewed (13), or HL-60 cells (14). Sphingosine has been shown to in Refs. 3- 5). activate PLC and diacylglycerol kinase in Jurkat T cells To date, evidence has accumulated that receptor-stimu (15) or airway smooth muscle (16). We have shown that lated activation of glycerophospholipid metabolism, which sphingosine enhances U46619- and thrombin-induced PLC is initiated by the activation of a variety of phospholipases, activation (17), and phosphatidylinositol 4-kinase activa e.g., phospholipase (PL) A2, C, and D, is linked to the tion (18) in rabbit platelets, suggesting the contribution of sphingosine to receptor-stimulated phosphatidylinositol 1This work was supported by a Grant-in-Aid for ScientificResearch turnover. Chao et al. also reported that sphingosine en (No. 10672072) from the Ministry of Education, Science,Sports and hances inositol phosphate accumulation by stimulating PLC Culture of Japan. activity in human foreskin fibroblasts (19) . 2To whom correspondenceshould be addressed. Fax: +81-75-595- On the other hand, the involvement of sphingolipids in 4759, E-mail: [email protected],jp cellular responses associated with PLA2 activation has been Abbreviations: C2-ceramide,N-acetylsphingosine; C6-ceramide,N- reported. C2-Ceramide and sphingosine have been shown to hexanoylsphingosine; ePLA2, group IV cytosolic phospholipase A2; DAG, diacylglycerol; MAPK, mitogen-activated protein kinase; stimulate interleukin-ƒÀ-induced prostaglandin E 2 genera PKC, protein kinase C; PL, phospholipase. tion concomitant with an increase in the induction of cyclooxygenase mRNA in human dermal fibroblasts (20) . c 1999 by The Japanese BiochemicalSociety. The C2- and C6-ceramides have also been reported to 96 J. Biochem . Potentiation of Phospholipase A2 by Ceramide 97 stimulate the expression of cyclooxygenase 2 and cPLA2 0.35% BSA, and 3.8mM HEPES, pH 6.5) containing 0.4 mRNAs in murine fibrosarcoma L929 cells (21). On the mM EGTA. The washed platelets were resuspended in contrary, it has been shown that arachidonic acid liberated HEPES buffer (pH 7.4). upon stimulation with tumor necrosis factor a mediates Lipid Metabolism-Platelet-rich plasma was incubated ligand-induced sphingomyelinase activation followed by with [3H] arachidonic acid (1ƒÊCi/ml) or [3H] glycerol (60 ceramide generation in HL-60 cells (22) and L929 murine ƒÊ Ci/ml) at 37°C for 1.5 h, and then washed as described fibroblast cell line (23), suggesting that PLA2 plays an above. The labeled platelets (5 x 108 cells/ml) were pre important role in the sphingolipid signaling pathway. treated with 100ƒÊM BW755C, a cyclooxygenase and Recently, we showed that sphingosine and C6-ceramide lipoxygenase inhibitor (30), at 37•Ž for 2 min in the enhance thrombin- and U46619-induced arachidonic acid presence of 1mM CaCl2. The platelets were further treated liberation in rabbit platelets (24, 25). In the present study, with C6-ceramide and then stimulated with various ago we extended our recent studies by defining the mechanism nists. When platelets were treated with PD98059 or by which ceramide enhances thrombin-stimulated ara U73122, the inhibitor was added before the treatment with chidonic acid liberation using cell-permeable C6-ceramide C6-ceramide. The reaction was terminated by adding in rabbit platelets. Although the activation mechanism is chloroform/methanol/HCl (200:200:1, v/v/v). Lipids well characterized in platelets, the step of receptor-stimu were extracted and separated by TLC on silica gel G plates lated arachidonic acid liberation followed by thromboxane with the following development systems: for the analysis of formation is a critical one for platelet activation. Several arachidonic acid and diacylglycerol, petroleum ether/dieth studies have demonstrated that the thrombin-stimulated yl ether/acetic acid (40:40:1, v/v/v); and for the analysis arachidonic acid liberation is caused by the catalytic action of lysophosphatidylcholine, chloroform/methanol/H20 of group IV cytosolic PLA2 (cPLA2) (26-28). Furthermore, (65:35:6, v/v/v). The area corresponding to each lipid was it has been reported that phorbol 12,13-dibutyrate, a scraped off and the radioactivity was determined by liquid scintillation counting. protein kinasss C (PKC) activator, phosphorylates mitogen - activated pppprotein kinase (MAPK), which in turn results in Measurement of the Activities of cPLA2, MAPK, and the phosphorylation of cPLA2 in platelets (29), suggesting PKC-Washed platelets (5•~108 cells/ml) were treated that cPLA2 activation is regulated, at least in part, by PKC with C6-ceramide in the presence of 1mM CaCl2, and then through MAPK. stimulated with thrombin. After centrifugation in the Therefore, in the present work we focused on the presence of 3mM EGTA, the platelets were washed and involvement of ceramide in thrombin-induced cPLA2 lysed by sonication in lysis buffer consisting of 100mM activation via the PLC-initiated sequential pathway includ NaCl, 20mM ƒÀ-glycerophosphate, 1mM Na3VO4, 2mM ing PKC and MAPK. Although platelets generate little EGTA, 10ƒÊM leupeptin, 100ƒÊM p-(amidinophenyl)- ceramide upon stimulation because of the low activity of methanesulfonyl fluoride, and 10mM Tris-HCl (pH 7.4). The lysate was centrifuged at 100,000 x g for 1 h at 4°C, sphingomyelinase, they seem to be more suitable for investigating the effect of ceramide, since in platelets the and the pellet was resuspended in the lysis buffer. The supernatant was subjected to assaying for the activity of effect of ceramide-induced signals to the nucleus on the enzyme activation can be excluded. cPLA2 or MAPK. The pellet was subjected to PKC assay- ing. The protein concentration of each sample was deter- mined with a commercial assay kit (Pierce, Rockford, IL). MATERIALS AND METHODS For the cPLA2 assay, the supernatant was treated with 5 Materials-C6-Ceramide (N-hexanoylsphingosine) and mM dithiothreitol to inhibit secretary PLA2 activity (31). cPLA2 activity in the supernatant was determined by phorbol 12-myristate 13-acetate were obtained from incubation of the supernatant with a mixture of 1-stearoyl- Sigma. A23187 and PD98059 (2'-amino- 3'-methoxyfla 2-[3H] arachidonoylglycerophosphocholine and the unlabel vone) were from Calbiochem (La Jolla, CA, USA). Throm ed compound (25 Ci/mol, 10ƒÊM) as a substrate at 37•Ž for bin (bovine plasma) was from Mochida Pharmaceuticals 5 min in the presence of 5mM CaCl2 and 100mM Tris-HC (Tokyo),
Load more

Recommended publications
  • Download Product Insert (PDF)
    Product Information U-46619 Item No. 16450 CAS Registry No.: 56985-40-1 Formal Name: 9,11-dideoxy-9α,11α-methanoepoxy-prosta- 5Z,13E-dien-1-oic acid Synonym: 9,11-dideoxy-9α,11α-methanoepoxy COOH Prostaglandin F2α MF: C21H34O4 O FW: 350.5 Purity: ≥98% OH Stability: ≥2 years at -20°C Supplied as: A solution in methyl acetate Laboratory Procedures For long term storage, we suggest that U-46619 be stored as supplied at -20°C. It will be stable for at least two years. U-46619 is supplied as a solution in methyl acetate. To change the solvent, simply evaporate the methyl acetate under a gentle stream of nitrogen and immediately add the solvent of choice. Solvents such as DMSO, ethanol, and dimethyl formamide purged with an inert gas can be used. The solubility of U-46619 in these solvents is aproximately 100 mg/ml. Further dilutions of the stock solution into aqueous buffers or isotonic saline should be made prior to performing biological experiments. Ensure that the residual amount of organic solvent is insignificant since organic solvents may have physiological effects at low concentrations. If an organic solvent-free solution of U-46619 is needed, it can be prepared by evaporating the methyl acetate and directly dissolving the neat oil in aqueous buffers. The solubility of U-46619 in PBS (pH 7.2) is approximately 2 mg/ml. We do not recommend storing the aqueous solution for more than one day. 1 U-46619 is a stable analog of the endoperoxide prostaglandin H2, and a TP receptor agonist.
    [Show full text]
  • Role of Proaggregatory and Antiaggregatory Prostaglandins in Hemostasis
    Role of proaggregatory and antiaggregatory prostaglandins in hemostasis. Studies with combined thromboxane synthase inhibition and thromboxane receptor antagonism. P Gresele, … , G Pieters, J Vermylen J Clin Invest. 1987;80(5):1435-1445. https://doi.org/10.1172/JCI113223. Research Article Thromboxane synthase inhibition can lead to two opposing effects: accumulation of proaggregatory cyclic endoperoxides and increased formation of antiaggregatory PGI2 and PGD2. The elimination of the effects of the cyclic endoperoxides by an endoperoxide-thromboxane A2 receptor antagonist should enhance the inhibition of hemostasis by thromboxane synthase blockers. We have carried out a series of double-blind, placebo-controlled, crossover studies in healthy volunteers to check if this hypothesis may be operative in vivo in man. In a first study, in 10 healthy male volunteers, the combined administration of the thromboxane receptor antagonist BM 13.177 and the thromboxane synthase inhibitor dazoxiben gave stronger inhibition of platelet aggregation and prolonged the bleeding time more than either drug alone. In a second study, in 10 different healthy male volunteers, complete inhibition of cyclooxygenase with indomethacin reduced the prolongation of the bleeding time by the combination BM 13.177 plus dazoxiben. In a third study, in five volunteers, selective cumulative inhibition of platelet TXA2 synthesis by low-dose aspirin inhibited platelet aggregation and prolonged the bleeding time less than the combination BM 13.177 plus dazoxiben. In vitro, in
    [Show full text]
  • Vasoactive Responses of U46619, PGF2 , Latanoprost, and Travoprost
    Vasoactive Responses of U46619, PGF2␣, Latanoprost, and Travoprost in Isolated Porcine Ciliary Arteries Ineta Vysniauskiene, Reto Allemann, Josef Flammer, and Ivan O. Haefliger PURPOSE. To compare the vasoactive properties of the prosta- these responses can be modulated by SQ 29548 (TP-receptor noids U46619 (thromboxane A2 analogue), prostaglandin F2␣ antagonist) or AL-8810 (FP-receptor antagonist). (PGF2␣), latanoprost free acid, and travoprost free acid in isolated porcine ciliary arteries. METHODS. In a myograph system (isometric force measure- MATERIAL AND METHODS ment), quiescent vessels were exposed (cumulatively) to U46619, PGF , latanoprost, or travoprost (0.1 nM–0.1 mM). 2␣ Vessels Preparation Experiments were also conducted in the presence of SQ 29548 (TP-receptor antagonist; 3–10 ␮M) or AL-8810 (FP-receptor Porcine eyes were obtained from an abattoir immediately after death. antagonist; 3–30 ␮M). Contractions were expressed as the In cold modified Krebs-Ringer solution (NaCl 118 mM, KCl 4.7 mM, percentage of 100 mM potassium chloride-induced contrac- CaCl2 2.5 mM, K2PO4 1.2 mM, MgSO4 1.2 mM, NaHCO3 25 mM, tions. glucose 11.1 mM, and EDTA 0.026 mM), ciliary arteries were dissected 5 ␮ RESULTS. In quiescent vessels, contractions (concentration–re- and cut into 2-mm segments. In an organ chamber, two 45- m Ϯ tungsten wires were passed through the vessel’s lumen and attached to sponse curves) induced by (0.1 mM) PGF2␣ (87.9% 3.5%), U46619 (66.7% Ϯ 4.1%), and latanoprost (62.9% Ϯ 3.6%) were a force transducer for isometric force measurements (Myo-Interface; JP more pronounced (P Յ 0.001) than those induced by tra- Trading, Aarhus, Denmark).
    [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]
  • Activation of the Murine EP3 Receptor for PGE2 Inhibits Camp Production and Promotes Platelet Aggregation
    Activation of the murine EP3 receptor for PGE2 inhibits cAMP production and promotes platelet aggregation Jean-Etienne Fabre, … , Thomas M. Coffman, Beverly H. Koller J Clin Invest. 2001;107(5):603-610. https://doi.org/10.1172/JCI10881. Article The importance of arachidonic acid metabolites (termed eicosanoids), particularly those derived from the COX-1 and COX-2 pathways (termed prostanoids), in platelet homeostasis has long been recognized. Thromboxane is a potent agonist, whereas prostacyclin is an inhibitor of platelet aggregation. In contrast, the effect of prostaglandin E2 (PGE2) on platelet aggregation varies significantly depending on its concentration. Low concentrations of PGE2 enhance platelet aggregation, whereas high PGE2 levels inhibit aggregation. The mechanism for this dual action of PGE2 is not clear. This study shows that among the four PGE2 receptors (EP1–EP4), activation of EP3 is sufficient to mediate the proaggregatory actions of low PGE2 concentration. In contrast, the prostacyclin receptor (IP) mediates the inhibitory effect of higher PGE2 concentrations. Furthermore, the relative activation of these two receptors, EP3 and IP, regulates the intracellular level of cAMP and in this way conditions the response of the platelet to aggregating agents. Consistent with these findings, loss of the EP3 receptor in a model of venous inflammation protects against formation of intravascular clots. Our results suggest that local production of PGE2 during an inflammatory process can modulate ensuing platelet responses. Find the latest version: https://jci.me/10881/pdf Activation of the murine EP3 receptor for PGE2 inhibits cAMP production and promotes platelet aggregation Jean-Etienne Fabre,1 MyTrang Nguyen,1 Krairek Athirakul,2 Kenneth Coggins,1 John D.
    [Show full text]
  • Prostaglandins and Other Lipid Mediators
    Prostaglandins & other Lipid Mediators 122 (2016) 18–27 Contents lists available at ScienceDirect Prostaglandins and Other Lipid Mediators Original research article In silico modelling of prostacyclin and other lipid mediators to nuclear receptors reveal novel thyroid hormone receptor antagonist properties ∗ Noelia Perez Diaz, Mire Zloh, Pryank Patel, Louise S. Mackenzie Life and Medical Sciences, University of Hertfordshire, Hatfield AL10 9AB, UK a r t i c l e i n f o a b s t r a c t Article history: Prostacyclin (PGI2) is a key mediator involved in cardiovascular homeostasis, acting predominantly on Received 10 August 2015 two receptor types; cell surface IP receptor and cytosolic peroxisome proliferator activated receptor Received in revised form (PPAR) ␤/␦. Having a very short half-life, direct methods to determine its long term effects on cells 19 November 2015 is difficult, and little is known of its interactions with nuclear receptors. Here we used computational Accepted 7 December 2015 chemistry methods to investigate the potential for PGI2, beraprost (IP receptor agonist), and GW0742 Available online 11 December 2015 ␤ ␦ (PPAR / agonist), to bind to nuclear receptors, confirmed with pharmacological methods. In silico screening predicted that PGI , beraprost, and GW0742 have the potential to bind to different Abbreviations: 2 nuclear receptors, in particular thyroid hormone ␤ receptor (TR␤) and thyroid hormone ␣ receptor (TR␣). MLS, MLS000389544 Docking analysis predicts a binding profile to residues thought to have allosteric control on the TR ligand NSAID, non-steroidal anti-inflammatory drug binding site. Luciferase reporter assays confirmed that beraprost and GW0742 display TR␤ and TR␣ −5 −6 × × PPAR␤/␦, peroxisome proliferator activated antagonistic properties; beraprost IC50 6.3 10 mol/L and GW0742 IC50 4.9 10 mol/L.
    [Show full text]
  • Role of Curcumin on Tumor Angiogenesis in Hepatocellular Carcinoma
    Naresuan University Journal 2008; 16(3): 239-254 239 Role of Curcumin on Tumor Angiogenesis in Hepatocellular Carcinoma Pornphrom Chintana Department of Physiology, Faculty of Medical Science, Naresuan University, Phitsanulok 65000, Thailand. Corresponding author. E-mail address: [email protected] (P. Chintana) Received 14 July 2008; accepted 24 December 2008 Summary Hepatocellular carcinoma (HCC) is a malignant tumor characterized by active neovascularization. Vascular endothelial growth factor (VEGF) is the most important angiogenic factor that regulates the HCC development. Overexpression of VEGF enhances the HCC tumor growth associated with increase of angiogenesis in the tumor, whereas suppression of VEGF attenuates the tumor growth. Similarly, the cyclooxygenase-2 (COX-2) expression stepwisely increases during hepatocarcinogenesis. Curcurmin has been shown to inhibit several angiogenic biomarkers including, VEGF and COX-2 expression. Therefore, curcumin could be used as a candidate for the combined drug treatment for HCC in the future. Keywords: Curcumin; Angiogenesis; Hepatocellular carcinoma; VEGF; COX-2 INTRODUCTION greater practical significance than non-selective cytotoxic therapies to control the tumor growth Hepatocellular carcinoma (HCC) is one of the and metastasis by targeting angiogenesis. Since, five most common cancers worldwide, with a many dietary and non-dietary phytochemicals do particularly high prevalence in Asian countries due not affect survival of normal cells and also possess to endemic hepatitis B virus infection (Parkin et al., anti-angiogenic as well as anti-tumorigenic 2001). In Thailand, it is the most common cause of activities, it could be a rationale approach to examine death in men (Vatanasapt et al., 2002). Surgery, their inhibitory effect on tumor angiogenesis.
    [Show full text]
  • G Protein-Coupled Receptors
    S.P.H. Alexander et al. The Concise Guide to PHARMACOLOGY 2015/16: G protein-coupled receptors. British Journal of Pharmacology (2015) 172, 5744–5869 THE CONCISE GUIDE TO PHARMACOLOGY 2015/16: G protein-coupled receptors Stephen PH Alexander1, Anthony P Davenport2, Eamonn Kelly3, Neil Marrion3, John A Peters4, Helen E Benson5, Elena Faccenda5, Adam J Pawson5, Joanna L Sharman5, Christopher Southan5, Jamie A Davies5 and CGTP Collaborators 1School of Biomedical Sciences, University of Nottingham Medical School, Nottingham, NG7 2UH, UK, 2Clinical Pharmacology Unit, University of Cambridge, Cambridge, CB2 0QQ, UK, 3School of Physiology and Pharmacology, University of Bristol, Bristol, BS8 1TD, UK, 4Neuroscience Division, Medical Education Institute, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY, UK, 5Centre for Integrative Physiology, University of Edinburgh, Edinburgh, EH8 9XD, UK Abstract The Concise Guide to PHARMACOLOGY 2015/16 provides concise overviews of the key properties of over 1750 human drug targets with their pharmacology, plus links to an open access knowledgebase of drug targets and their ligands (www.guidetopharmacology.org), which provides more detailed views of target and ligand properties. The full contents can be found at http://onlinelibrary.wiley.com/doi/ 10.1111/bph.13348/full. G protein-coupled receptors are one of the eight major pharmacological targets into which the Guide is divided, with the others being: ligand-gated ion channels, voltage-gated ion channels, other ion channels, nuclear hormone receptors, catalytic receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8.470,805 B2 Chen (45) Date of Patent: Jun
    USOO8470805B2 (12) United States Patent (10) Patent No.: US 8.470,805 B2 Chen (45) Date of Patent: Jun. 25, 2013 (54) PROCESSES FOR PREPARING OTHER PUBLICATIONS PPERAZINUM SALTS OF KMUP AND USE THEREOF Arayne et al. Med Chem Res. (2010) 19:717-731.* Chung et al., “The xanthine derivative KMUP-1 inhibits models of pulmonary artery hypertension via increased NO and coMP-depen (75) Inventor: Ing-Jun Chen, Kaohsiung (TW) dent inhibition of RhoA, Rho kinase.' British Journal of Pharmacol (73) Assignee: Kaohsiung Medical University, ogy, (Jun. 2010) 160(4):971-986. Evans et al., Effects of HMG-CoA Reductase Inhibitors on Skeletal Kaohsiung (TW) Muscle: Are all Statins the Same?, Drug Safety (2002) 25(9):649 663. (*) Notice: Subject to any disclaimer, the term of this Hiro et al., Effects of Intensive Statin Therapy of Regression of patent is extended or adjusted under 35 coronary Atherosclerosis in Patients with Acute Coronary Syndrome: U.S.C. 154(b) by 193 days. A Multicenter Randomized Trial Evaluated by Volumetric Intravascular Ultrasound Using Pitavastatin Versus Atorvastatin (21) Appl. No.: 12/878,451 (Japan-ACSJapan Assessment of Pitavastatin and Atorvastatin in Acute Coronary Syndrome Study), J. Am. Coll. Cardiol. (2009) (22) Filed: Sep. 9, 2010 54:293-302. Jacobson TA, “Myopathy with statin-fibrate combination therapy: (65) Prior Publication Data clinical considerations.” Nat. Rev. endocrinol. (Sep. 2009) 5(9):507 US 2011 FO136767 A1 Jun. 9, 2011 518. Lin et al., “KMUP-1 relaxes rabbit corpus cavernosum smooth muscle in vitro and in vivo: involvement of cyclic GMP and K" Related U.S.
    [Show full text]
  • Supplementary Information Development of Fatty Acid
    JPET#234781 Supplementary Information Development of fatty acid derivatives of aspirin for inhibition of platelet aggregation Jahnabi Roy c, Reheman Adili e, Richard Kulmacz f, Michael Holinstat e,g, and Aditi Das* a, b, d a Department of Comparative Biosciences, b Department of Biochemistry, Center for Biophysics and Quantitative Biology, Division of Nutritional Sciences, Department of Bioengineering c Department of Chemistry d Beckman Institute for Advanced Science, f The Department of Internal Medicine, Texas Health Science Center at Houston, McGovern Medical School, Houston, TX 77225-0708, e Department of Pharmacology, g Division of Cardiovascular Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, c, a University of Illinois Urbana-Champaign, Urbana IL 61801. Journal: THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS (JPET) CONTENTS Fig S1: Inhibition of platelet aggregation by various inducers S2 Fig S2: Inhibition of platelet aggregation by various inducers -2 S3 Fig S3: TXB2 formed in platelet aggregometry samples S4 Fig S4: Inhibition of platelet aggregation by LA, EPA and DHA S5 Fig S5: Coupled assay schematic and results for parent fatty acids S6 Fig S6: Coupled assay for LA, EPA, DHA and aspirin S7 Fig S7: Representative HPLC trace of DHA anhydride hydrolysis S8 Fig S8: Representative HPLC trace of EPA anhydride hydrolysis S9 Fig S9: Representative HPLC trace of LA anhydride hydrolysis S10 Table 1: NMR and MS characterization of compounds S11 Fig S10: NMR of DHA anhydride S12 Fig S11: NMR of EPA anhydride S13 Fig S12: NMR of LA anhydride S14 S1 JPET#234781 Supplemental figure 1: Inhibition of platelet aggregation in washed platelets by anhydrides (10 μM) in the presence of (A) 1nM Thrombin (B) 50 μM PAR4-AP (C) 1 μM PAR1-AP (D) 1 μM ADP (E) 1 μM U46619 (thromboxane A2 analog).
    [Show full text]
  • Ginsenoside-Rd Induces Cholesterol Efflux from Macrophage-Derived Foam Cells Conjunction with Cardiovascular, Pharmacological and Biochemical Evaluations
    npg Section 6: Cardiovascular Pharmacology 56 10.1038/aps.2013.113 model of brain stem death that employed microinjection of the organophosphate S6.1 insecticide mevinphos bilaterally into RVLM of Sprague-Dawley rats was used, in Ginsenoside-Rd induces cholesterol efflux from macrophage-derived foam cells conjunction with cardiovascular, pharmacological and biochemical evaluations. Da-yan CAO, Ya LIU*, Xiao-hui LI*. Institute of Materia Medica and Department of Results: A significant increase in TrkB protein, phosphorylation of TrkB at Tyr516 Pharmaceutics, College of Pharmacy, Third Military Medical University, Chongqing (pTrkBY516), Shc at Tyr317 (pShcY317) or extracellular signal-regulated kinase (ERK) 400038, China at Thr202/Tyr204, or Ras activity in RVLM occurred preferentially during the pro- Aim: Ginsenoside-Rd, a purified component from panax notoginseng saponins, life phase of experimental brain stem death. Microinjection bilaterally into RVLM has been described to reduce atherosclerotic lesion formation, yet the mechanism of a specific TrkB inhibitor, K252a, antagonized those increases. Pretreatment with is not fully understood. This study is designed to investigate the potential role of anti-pShcY317 antiserum, Src homology 3 binding peptide (Grb2/SOS inhibitor), Ginsenoside-Rd in modulating cholesterol deposition and to explore its underlying farnesylthioacetic acid (Ras inhibitor), manumycin A (Ras inhibitor) or GW5074 mechanisms in macrophages. Methods: The murine macrophage-derived foam (Raf-1 inhibitor) blunted the preferential augmentation of Ras activity or ERK cells induced by 25 μg/mL oxidized low-density lipoprotein (oxLDL) in RAW264.7 phosphorylation in RVLM and blocked the upregulated nitric oxide synthase I cells for 48 h. Ginsenoside-Rd 10, 20 μmol/L was added during oxLDL incubation (NOS I)/protein kinase G (PKG) signaling, the pro-life cascade that sustains central respectively.
    [Show full text]
  • Oxidative Stress Inducers Potentiate 2, 3, 7, 8-Tetrachlorodibenzo-P-Dioxin
    Advance Publication The Journal of Veterinary Medical Science Accepted Date: 10 May 2021 J-STAGE Advance Published Date: 24 May 2021 ©2021 The Japanese Society of Veterinary Science Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. TOXICOLOGY FULL PAPER Oxidative stress inducers potentiate 2,3,7,8-tetrachlorodibenzo-p- dioxin-mediated pre-cardiac edema in larval zebrafish Katsuki Tanaka 1), Hikaru Adachi 1), Hironobu Akasaka 1), Junya Tamaoki 2), Yuji Fuse 2), Makoto Kobayashi 2), Takio Kitazawa 1) and Hiroki Teraoka 1)* 1) School of Veterinary Medicine, Rakuno Gakuen University, 069-8501 Ebetsu, Japan 2) Department of Molecular and Developmental Biology, Faculty of Medicine, University of Tsukuba, 305-8575 Tsukuba, Japan Running Head: Oxidative stress in TCDD toxicity ∗Corresponding author at: School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501, Japan. Tel.: +81 11 388 4791;fax: +81 11 388 4791. E-mail address: [email protected] (H. Teraoka). ABSTRACT We reported the involvement of oxidative stress and prostaglandins including thromboxane and prostacyclin in pre-cardiac edema (early edema) caused by 2,3,7,8- tetrachlorodibenzo-p-dioxin (TCDD). While the involvement of oxidative stress in TCDD-induced toxicity has been frequently reported, the mechanism of its action is still unclear. In the present study, oxidative stress inducers including paraquat, hydrogen peroxide (H2O2) and rotenone augmented early edema (edema) induced by a low concentration of TCDD (0.1 ppb) at 55 hours post fertilization (hpf), while each of them alone did not cause edema. Edema caused by TCDD plus oxidative stress inducers was almost abolished by antioxidants, an antagonist for thromboxane receptor (ICI-192,605) and an agonist for prostacyclin receptor (beraprost), suggesting that the site of action of these inducers was in the regular signaling pathway after activation of aryl hydrocarbon receptor type 2 (AHR2) by TCDD.
    [Show full text]