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purinergic 2X7Receptor is Involved in the Podocyte Damage of Obesity-Related Glomerulopathy via Activating Nucleotide-Binding and Oligomerization Domain-Like Receptor Protein 3 Inflammasome

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Chinese Medical Journal 2018年第22期131卷 2713-2725页

作者： Xiao-Xia Hou Hong-Rui Dong Li-Jun Sun Min Yang Hong Cheng Yi-Pu Chen Division of Nephrology Beijing Anzhen HospitalCapital Medical UniversityBeijing 100029China

Background:The nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome composed of NLRP3,apoptosis-associated speck-like protein containing CARD (ASC),and caspase-1 is engaged in the inflammatory response of many kidney diseases and can be activated by purinergic 2X7 receptor (P2X7R).This study was conducted to explore whether P2X7R plays a pathogenic role in the podocyte damage of obesity-related glomerulopathy (ORG) and whether this role is mediated by the activation ofNLRP3 inflammasome.Methods:A mouse model of ORG was established by high-fat diet feeding.The conditionally immortalized mouse podocytes were cultured with leptin or with leptin and P2X7R antagonist (KN-62 or A438079).The mRNA and protein expression of the P2X7R and NLRP3 inflammasome components including NLRP3,ASC,and caspase-1,as well as the podocyte-associated molecules including nephrin,podocin,and desmin in mouse renal cortex or cultured mouse podocytes were tested by real-time-polymerase chain reaction and Westem blot analysis,respectively.Results:The significantly upregulated expression of P2X7R and NLRP3 inflammasome components and the NLRP3 inflammasome activation were observed in the renal cortex (in fact their location in podocytes was proved by confocal microscopy) of ORG mice in vivo,which were accompanied with the morphological changes of podocyte damage and the expression changes of podocyte-associated molecules.Similar changes in the expression of P2X7R and NLRP3 inflammasome components as well as in the expression ofpodocyte-associated molecules were also observed in the cultured podocyte studies treated by leptin in vitro,and all of the above changes were significantly attenuated by the P2X7R antagonist KN-62 or A438079.Conclusions:P2X7R could trigger the activation ofNLRP3 inflammasome,and the activated P2X7R/NLRP3 inflammasome in podocytes might be involved in the podocyte damage of ORG.

关键词： Desmin Nucleotide-Binding and Oligomerization Domain-Like Receptor Protein 3 Inflammasome Obesity-Related Glomerulopathy Podocytes purinergic 2X7Receptor

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Autoimmune Hepatitis: Clinical Review with Insights into the purinergic Mechanism of Disease

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Journal of Clinical and Translational Hepatology 2013年第2期1卷 79-86页

作者： Nikhil Kapila Jennifer T.Higa Maria Serena Longhi Simon C.Robson Department of Medicine University of Connecticut Farmington CT USA Gastroenterology Division and Department of Medicine Beth Israel Deaconess Medical Center and Harvard Medical School Boston MA USA Gastroenterology Division and Department of Medicine Beth Israel Deaconess Medical Center and Harvard Medical School Boston MA USA Institute of Liver Studies King's College London School of Medicine at King's College Hospital Denmark Hill London UK

Autoimmune hepatitis (AIH) is an important disorder that predominantly results in inflammatory liver disease in genetically predisposed women.The clinicopathological picture is characterized by symptoms associated with both systemic inflammation and hepatic dysfunction,and with increased serum aminotransferases,elevated IgG,autoantibodies,and interface hepatitis on liver biopsy.AIH usually results in liver injury as a consequence of chronic hepatitis and cirrhosis.However,rarely,patients may present with fulminant liver failure.Early diagnosis is important in all instances because the disease can be highly responsive to immunosuppressive therapeutic options.Left untreated,the disease is associated with high morbidity and mortality.Here we provide an overview of the current state of knowledge on AIH and summarize the treatment options for this serious condition in adults.We also discuss the pathogenesis of the disease as a possible consequence of autoimmunity and the breakdown of hepatic tolerance.We focus on regulatory T cell impairments as a consequence of changes in CD39 ectonucleotidase expression and altered purinergic signaling.Further understanding of hepatic tolerance may aid in the development of specific and well-tolerated therapies for AIH.

关键词： Autoimmune hepatitis Pathogenesis Immunology purinergic CD39 Therapeutic overview

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WJSC 6^(th) Anniversary Special Issues(2):Mesenchymal stem cells purinergic receptors and nucleotide processing ectoenzymes:Their roles in regulating mesenchymal stem cell functions

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World Journal of Stem Cells 2014年第2期6卷 153-162页

作者： Sonia Scarfì Department of Earth Environment and Life SciencesUniversity of Genova

Human mesenchymal stem cells(MSCs)are a rare population of non-hematopoietic stem cells with multilineage potential,originally identified in the bone marrow.Due to the lack of a single specific marker,MSCs can be recognized and isolated by a series of features such as plastic adherence,a panel of surface markers,the clonogenic and the differentiation abilities.The recognized role of MSCs in the regulation of hemopoiesis,in cell-degeneration protection and in the homeostasis of mesodermal tissues through their differentiation properties,justifies the current interest in identifying the biochemical signals produced by MSCs and their active crosstalk in tissue environments.Only recently have extracellular nucleotides(eNTPs)and their metabolites been included among the molecular signals produced by MSCs.These molecules are active on both ionotropic and metabotropic receptors present in most cell types.MSCs possess a significant display of these receptors and of nucleotide processing ectoenzymes on their plasma membrane.Thus,from their niche,MSCs give a significant contribution to the complex signaling network of eNTPs and its derivatives.Recent studies have demonstrated the multifaceted aspects of eNTP metabolism and their signal transduction in MSCs and revealed important roles in specifying differentiation lineages and modulating MSC physiology and communication with other cells.This review discusses the roles of eNTPs,their receptors and ectoenzymes,and the relevance of the signaling network and MSC functions,and also focuses on the importance of this emerging area of interest for future MSC-based cell therapies.

关键词： Mesenchymal stem cell purinergic receptors Ectoenzymes ATP β-NAD Adenosine cADPR

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Genomic Organization of purinergic P2X Receptors

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Pharmacology & Pharmacy 2015年第8期6卷 341-362页

作者： Raúl Loera-Valencia Josué Obed Jaramillo-Polanco Andrómeda Linan-Rico María Guadalupe Nieto Pescador Juan Francisco Jiménez Bremont Carlos Barajas-López División de Biología Molecular Instituto Potosino de Investigación Científica y Tecnológica San Luís Potosí México

purinergic P2X receptors are a family of ligand-gated cationic channels activated by extracellular ATP. P2X subunit protein sequences are highly conserved between vertebrate species. However, they can generate a great diversity of coding splicing variants to fulfill several roles in mammalian physiology. Despite intensive research in P2X expression in both central and peripheral nervous system, there is little information about their homology, genomic structure and other key features that can help to develop selective drugs or regulatory strategies of pharmacological value which are lacking today. In order to obtain clues on mammalian P2X diversity, we have performed a bioinformatics analysis of the coding regions and introns of the seven P2X subunits present in human, simian, dog, mouse, rat and zebrafish. Here we report the arrangements of exon and intron sequences, considering its number, size, phase and placement;proposing some ideas about the gain and loss of exons and retention of introns. Taken together, these evidences show traits that can be used to gain insight into the evolutionary history of vertebrate P2X receptors and better understand the diversity of subunits coding the purinergic signaling in mammals.

关键词： Alternative Splicing Intron Genomic Organization P2X purinergic Signalling

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Uridine adenosine tetraphosphate acts as a pro-angiogenic factor in vitrothrough purinergic P2Y receptors

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中国药理学与毒理学杂志 2015年第S1期29卷 114-114页

作者： Zhi-chaoZHOU IhsanCHRIFI Yan-juanXU DirkJDUNCKER SJamalMUSTAFA DaphneMERKUS CarolineCHENG Division of Experimental Cardiology Cardiovascular Research School COEUR Erasmus MC University Medical Center Rotterdam Molecular Cardiology Department of Cardiology Thoraxcenter Laboratory of Renal and Vascular Biology Department of Nephrology &Hypertension University Medical Center Utrecht Department of Physiology and Pharmacology Center for Cardiovascular and Respiratory Sciences West Virginia University

OBJECTIVE Uridine adenosine tetraphosphate(Up4A),a dinucleotide,contains both purine and pyrimidine moieties,and exerts its vascular influence via activation of purinergic receptors.Here,we aimed to investigate the effects of Up4 A on angiogenesis and the putative purinergic receptors(PR)involved in this process.METHODS Tubule formation assay was performed in 3D matrix system.In this assay,human umbilical vein endothelial cells(HUVECs)were co-cultured with pericytes with various Up4 A doses(0,1,2.5,5,10 and 20μmol·L-1)in the absence and presence of P2Y6 R antagonist MRS2578(10μmol·L-1)for 5d.Expression profile of PR subtypes and angiogenic factors was assessed in HUVECs by q-PCR with and without P2Y6 R antagonist.RESULTS No difference in initial tubule formation was detected between Up4 A stimulation and control conditions at day 2.In contrast,a significant increase in vascular density in response to Up4 A was observed at day 5.Up4 A at a dose of 2.5and 5μmol·L-1 promoted total tubule length(by-1.89 fold and-2.23fold),number of tubules(by-1.71 fold and-1.89fold)as well as number of junctions(by-2.24 fold and-2.80fold),all of which were inhibited by MRS2578.Further increase in Up4 A dose to10 and 20μmol·L-1 did not induce an increase in these vascular parameters as compared to non-treated controls.Moreover,Up4 A increased mRNA level of P2YRs(P2Y2R,P2Y4 R and P2Y6R)but not P2XR(P2X4R and P2X7R)or P1R(A2AR and A2BR),while Up4 A upregulated VEGFA and ANGPT1 but not VEGFR2,ANGPT2,Tie1 and Tie2at mRNA level.Transcriptional upregulation of P2 YRs and angiogenic factors by Up4 A was inhibited by MRS2578.CONCLUSION Up4 A is functionally capable of promoting tubule formation in vitro co-culture system.This process is likely mediated by activation of pyrimidine-favored P2 YRs but not P2 XR or P1 Rs,and involves stimulation of well known angiogenic factors.

关键词： Up4A purinergic receptors angiogenesis P2Y6 tubule

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P2X7 receptor signaling during adult hippocampal neurogenesis

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Neural Regeneration Research 2019年第10期14卷 1684-1694页

作者： Hannah C. Leeson Tailoi Chan-Ling Michael D. Lovelace Jeremy C. Brownlie Ben J. Gu Michael W. Weible II Australian Institute for Bioengineering and Nanotechnology The University of QueenslandBrisbaneQueenslandAustralia Griffith Institute for Drug Discovery Griffith UniversityBrisbaneQueenslandAustralia Discipline of Anatomy and Histology School of Medical ScienceThe University of SydneySydneyNew South WalesAustralia Bosch Institute The University of SydneySydneyNew South WalesAustralia Applied Neurosciences Program Peter Duncan Neurosciences Research UnitSt. Vincent’s Centre for Applied Medical ResearchSydneyNew South WalesAustralia Faculty of Medicine St. Vincent’s Clinical SchoolUniversity of New South Wales SydneySydneyNew South WalesAustralia School of Environment and Science Griffith UniversityBrisbaneQueenslandAustralia Florey Institute of Neuroscience and Mental Health University of MelbourneMelbourneVictoriaAustralia

Neurogenesis is a persistent and essential feature of the adult mammalian hippocampus.Granular neurons generated from resident pools of stem or progenitor cells provide a mechanism for the formation and consolidation of new memories.Regulation of hippocampal neurogenesis is complex and multifaceted,and numerous signaling pathways converge to modulate cell proliferation,apoptosis,and clearance of cellular debris,as well as synaptic integration of newborn immature neurons.The expression of functional P2X7 receptors in the central nervous system has attracted much interest and the regulatory role of this purinergic receptor during adult neurogenesis has only recently begun to be explored.P2X7 receptors are exceptionally versatile:in their canonical role they act as adenosine triphosphate-gated calcium channels and facilitate calcium-signaling cascades exerting control over the cell via calcium-encoded sensory proteins and transcription factor activation.P2X7 also mediates transmembrane pore formation to regulate cytokine release and facilitate extracellular communication,and when persistently stimulated by high extracellular adenosine triphosphate levels large P2X7 pores form,which induce apoptotic cell death through cytosolic ion dysregulation.Lastly,as a scavenger receptor P2X7 directly facilitates phagocytosis of the cellular debris that arises during neurogenesis,as well as during some disease states.Understanding how P2X7 receptors regulate the physiology of stem and progenitor cells in the adult hippocampus is an important step towards developing useful therapeutic models for regenerative medicine.This review considers the relevant aspects of adult hippocampal neurogenesis and explores how P2X7 receptor activity may influence the molecular physiology of the hippocampus,and neural stem and progenitor cells.

关键词： P2X7 P2X7R adult neurogenesis neural stem cells neural progenitor cells hippocampus SGZ calcium signaling purinergic signaling

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purinergic signaling and human immunodeficiency virus/acquired immune deficiency syndrome: From viral entry to therapy

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World Journal of Virology 2015年第3期4卷 285-294页

作者： Daniela F Passos Maria Rosa C Schetinger Daniela BR Leal Departamento de Microbiologia e Parasitologia Centro de Ciências da SaúdeUniversidade Federal de Santa Maria Departamento de Bioquímica e Biologia Molecular Centro de Ciências Naturais e ExatasUniversidade Federal de Santa Maria

Human immunodeficiency virus(HIV) infection is a serious condition associated to severe immune dysfunction and immunodeficiency. Mechanisms involved in HIV-associated immune activation, inflammation and loss of CD4+ T cells have been extensively studied, including those concerning purinergic signaling pathways. purinergic signaling components are involved in viral entry and replication and disease progression. Research involving the participation of purinergic signaling in HIV infection has been not only important to elucidate disease mechanisms but also to introduce new approaches to therapy. The involvement of purinergic signaling in the pathogenesis of HIV infection and its implications in the control of the HIV infection are reviewed in this paper.

关键词： Human immunodeficiency virus purinergic signaling Immune activation Inflammation

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purinergic signaling in the gastrointestinal tract

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World Journal of Gastrointestinal Pathophysiology 2011年第2期2卷 31-34页

作者： Geoffrey Burnstock Autonomic Neuroscience Centre University College Medical SchoolRoyal Free Campus

Geoffrey Burnstock completed a BSc at King's College London and a PhD at University College London. He held postdoctoral fellowships with Wilhelm Feldberg(National Institute for Medical Research),Edith Bülbring(University of Oxford) and C. Ladd Prosser(University of Illinois) . He was appointed to a Senior Lectureship in Melbourne University in 1959 and became Professor and Chairman of Zoology in 1964. In 1975 he became Head of Department of Anatomy and Developmental Biology at UCL and Convenor of the Center of Neuroscience. He has been Director of the Autonomic Neuroscience Institute at the Royal Free Hospital School of Medicine since 1997. He was elected to the Australian Academy of Sciences in 1971,the Royal Society in 1986,the Academy of Medical Sciences in 1998 and an Honorary Fellow of the Royal College of Surgeons and the Royal College of Physicians in 1999 and 2000. He was awarded the Royal Society Gold Medal in 2000. He is editor-in-chief of the journals Autonomic Neuroscience and purinergic Signalling and on the editorial boards of many other journals. Geoffrey Burnstock's major research interest has been autonomic neurotransmission and he is best known for his seminal discovery of purinergic transmission and receptors,their signaling pathways and functional relevance. He has supervised over 100 PhD and MD studentsand published over 1400 original papers,re-views and books. He was first in the Institute of Scientific Information list of most cited scientists in Pharmacology and Toxicology from 1994-2004 [59.083 citations(March 2011) and an h-index of 109].

关键词： Geoffrey Burnstock purinergic signaling Gastrointestinal tract

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Effect of clopidogrel in bone healing-experimental study in rabbits

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World Journal of Orthopedics 2019年第12期10卷 434-445页

作者： Theodoros Lillis Alexander Veis Nikolaos Sakellaridis Anastasios Tsirlis Zoe Dailiana Department of Orthopaedic Surgery Faculty of MedicineUniversity of ThessalyBiopolisLarissa 41500Greece Department of Dentoalveolar Surgery Implantology and Oral RadiologyFaculty of DentistryAristotle University of ThessalonikiPanepistimioupoliThessaloniki 54124Greece Department of Clinical Pharmacology Faculty of MedicineUniversity of ThessalyBiopolisLarissa 41500Greece

BACKGROUND Clopidogrel is a widely prescribed drug for prevention of myocardial infarction and stroke in patients at risk.It inhibits thrombus formation via inhibition of the P2Y12 purinergic receptor on platelets,which is important in their activation by ADP.However,the P2Y12 receptor has also been found to be expressed in both osteoblasts and osteoclasts.Accumulated evidence suggests that purinergic receptors regulate important functions of bone turnover.Previous studies on the effect of clopidogrel on bone metabolism indicated potential harmful effects,but their results remain conflicting.Thus,clopidogrel treatment may affect bone healing,but it has not yet been studied.AIM To evaluate if continuous perioperative clopidogrel treatment has any negative effect on bone healing in the rabbit calvarial defect model.METHODS Sixteen male white New Zealand rabbits were randomly assigned in two groups:One group received daily 3 mg/kg of clopidogrel per os and the other group received the vehicle alone for a week prior to the surgical procedures;the treatments were continued for another 6 wk postoperatively.The surgical procedures included generation of two circular calvarial defects 11 mm in diameter in every animal.After the 6-wk period of healing,postmortem radiographic and histomorphometric evaluation of the defects was performed.RESULTS Both the surgical procedures and the postoperative period were uneventful and well tolerated by all the animals,without any surgical wound dehiscence,signs of infection or other complication.New bone was formed either inwards from the defect margins or in the central portion of the defect as separated bony islets.While defect healing was still incomplete in both groups,the clopidogrel group had significantly improved radiographic healing scores.Moreover,the histomorphometric analysis showed that bone regeneration(%)was 28.07±7.7 for the clopidogrel group and 19.47±4.9 for the control group,showing a statistically significant difference between them(P=0.018).Statistically significant

关键词： Clopidogrel Bone healing purinergic signaling Calvarial defect Rabbit

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P2X7 receptor activation causes phosphatidylserine exposure in canine erythrocytes

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World Journal of Hematology 2016年第4期5卷 88-93页

作者： Megan Faulks Tracey A Kuit Reece A Sophocleous Belinda L Curtis Stephen J Curtis Lisa M Jurak Ronald Sluyter School of Biological Sciences University of Wollongong Centre for Medical and Molecular Bioscience University of Wollongong Illawarra Health and Medical Research Institute Albion Park Veterinary Hospital

AIM To determine if activation of the ATP-gated P2X7 receptor channel induces phosphatidylserine(PS) exposure in erythrocytes from multiple dog breeds.METHODS Peripheral blood was collected from 25 dogs representing 13 pedigrees and seven crossbreeds. ATP-induced PS exposure on canine erythrocytes in vitro was assessed using a flow cytometric Annexin V binding assay.RESULTS ATP induced PS exposure in erythrocytes from all dogs studied. ATP caused PS exposure in a concentrationdependent manner with an EC50 value of 395 μmol/L. The non-P2X7 agonists, ADP or AMP, did not cause PS exposure. The P2X7 antagonist, AZ10606120, but not the P2X1 antagonist, NF449, blocked ATP-induced PS exposure.CONCLUSION The results indicate that ATP induces PS exposure in erythrocytes from various dog breeds and that this process is mediated by P2X7 activation.

关键词： Adenosine triphosphate Dog P2X1 receptor P2X7 receptor Phospholipid purinergic receptor Red blood cells

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