Short Exposure to Ethanol Diminishes Caspase-1 and ASC Activation in Human HepG2 Cells In Vitro

被引:15
作者
Hoerauf, Jason-Alexander [1 ]
Kany, Shinwan [2 ,3 ]
Janicova, Andrea [2 ]
Xu, Baolin [2 ]
Vrdoljak, Teodora [4 ]
Sturm, Ramona [1 ]
Dunay, Ildiko Rita [5 ]
Martin, Lukas [6 ]
Relja, Borna [2 ]
机构
[1] Goethe Univ Frankfurt, Dept Trauma Hand & Reconstruct Surg, D-60438 Frankfurt, Germany
[2] Otto von Guericke Univ, Dept Radiol & Nucl Med, Expt Radiol, D-39108 Magdeburg, Germany
[3] Univ Hosp Hamburg Eppendorf, Univ Heart Ctr, Dept Cardiol Emphasis Electrophysiol, D-20251 Hamburg, Germany
[4] Univ Zagreb, Univ Hosp Dubrava, Dept Diagnost & Intervent Radiol, Sch Med, HR-10000 Zagreb, Croatia
[5] Otto von Guericke Univ, Inst Inflammat & Neurodegenerat, D-39108 Magdeburg, Germany
[6] Univ Hosp RWTH Aachen, Dept Intens Care & Intermediate Care, D-52074 Aachen, Germany
关键词
inflammation; inflammasome; caspase-1; alcohol; liver; in vitro; NLRP3 INFLAMMASOME ACTIVATION; LUNG EPITHELIAL-CELLS; NALP3; INFLAMMASOME; ALCOHOL; RESPONSES; LIVER; PHOSPHORYLATION; CONSUMPTION; EXPRESSION; CYTOKINES;
D O I
10.3390/ijms21093196
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
This paper discusses how the assembly of pro-caspase-1 and apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) in macromolecular protein complexes, inflammasomes, activates caspase-1. The present study investigates the molecular mechanisms of inflammasome activation in HepG2 cells and examines how short exposures to ethanol (EtOH) affect inflammasome activation. HepG2 cells were treated with lipopolysaccharide (LPS), ATP or nigericin (NIG) in a two-step model. After LPS priming, ATP or NIG were added. As inhibitors, sodium orthovanadate (general inhibitor of tyrosine phosphatases), AC-YVAD-CMK (caspase-1 inhibitor) or AZ10606120 (purinergic receptor P2X7R inhibitor) were applied after LPS priming. To monitor the inflammasome activation, the caspase-1 activity, ASC speck formation, reactive oxygen species (ROS) production and cell death were analyzed. To elucidate the mechanistical approach of EtOH to the inflammasome assembly, the cells were treated with EtOH either under simultaneous LPS administration or concurrently with ATP or NIG application. The co-stimulation with LPS and ATP induced a significant ASC speck formation, caspase-1 activation, cell death and ROS generation. The inhibition of the ATP-dependent purinoreceptor P2X7 decreased the caspase-1 activation, whereas sodium orthovanadate significantly induced caspase-1. Additional treatment with EtOH reversed the LPS and ATP-induced caspase-1 activation, ASC speck formation and ROS production. The ASC speck formation and caspase-1 induction require a two-step signaling with LPS and ATP in HepG2 cells. Inflammasome activation may depend on P2X7. The molecular pathway of an acute effect of EtOH on inflammasomes may involve a reduction in ROS generation, which in turn may increase the activity of tyrosine phosphatases.
引用
收藏
页数:18
相关论文
共 51 条
[1]   Tyrosine dephosphorylation and ethanol inhibition of N-methyl-D-aspartate receptor function [J].
Alvestad, RM ;
Grosshans, DR ;
Coultrap, SJ ;
Nakazawa, T ;
Yamamoto, T ;
Browning, MD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (13) :11020-11025
[2]  
[Anonymous], 2014, MEDIAT INFLAMM
[3]   Rutin modulates ASC expression in NLRP3 inflammasome: a study in alcohol and cerulein-induced rat model of pancreatitis [J].
Aruna, Ravikumar ;
Geetha, Arumugam ;
Suguna, Periyanayagam .
MOLECULAR AND CELLULAR BIOCHEMISTRY, 2014, 396 (1-2) :269-280
[4]   Inflammasome biology, molecular pathology and therapeutic implications [J].
Awad, Fawaz ;
Assrawi, Eman ;
Louvrier, Camille ;
Jumeau, Claire ;
Georgin-Lavialle, Sophie ;
Grateau, Gilles ;
Amselem, Serge ;
Giurgea, Irina ;
Karabina, Sonia-Athina .
PHARMACOLOGY & THERAPEUTICS, 2018, 187 :133-149
[5]   Boron-Based Inhibitors of the NLRP3 Inflammasome [J].
Baldwin, Alex G. ;
Rivers-Auty, Jack ;
Daniels, Michael J. D. ;
White, Claire S. ;
Schwalbe, Carl H. ;
Schilling, Tom ;
Hammadi, Halah ;
Jaiyong, Panichakorn ;
Spencer, Nicholas G. ;
England, Hazel ;
Luheshi, Nadia M. ;
Kadirvel, Manikandan ;
Lawrence, Catherine B. ;
Rothwell, Nancy J. ;
Harte, Michael K. ;
Bryce, Richard A. ;
Allan, Stuart M. ;
Eder, Claudia ;
Freeman, Sally ;
Brough, David .
CELL CHEMICAL BIOLOGY, 2017, 24 (11) :1321-+
[6]   The NLRP3 inflammasome is released as a particulate danger signal that amplifies the inflammatory response [J].
Baroja-Mazo, Alberto ;
Martin-Sanchez, Fatima ;
Gomez, Ana I. ;
Martinez, Carlos M. ;
Amores-Iniesta, Joaquin ;
Compan, Vincent ;
Barbera-Cremades, Maria ;
Yaguee, Jordi ;
Ruiz-Ortiz, Estibaliz ;
Anton, Jordi ;
Bujan, Segundo ;
Couillin, Isabelle ;
Brough, David ;
Arostegui, Juan I. ;
Pelegrin, Pablo .
NATURE IMMUNOLOGY, 2014, 15 (08) :738-+
[7]   Cutting Edge: NF-κB Activating Pattern Recognition and Cytokine Receptors License NLRP3 Inflammasome Activation by Regulating NLRP3 Expression [J].
Bauernfeind, Franz G. ;
Horvath, Gabor ;
Stutz, Andrea ;
Alnemri, Emad S. ;
MacDonald, Kelly ;
Speert, David ;
Fernandes-Alnemri, Teresa ;
Wu, Jianghong ;
Monks, Brian G. ;
Fitzgerald, Katherine A. ;
Hornung, Veit ;
Latz, Eicke .
JOURNAL OF IMMUNOLOGY, 2009, 183 (02) :787-791
[8]   Salmonella induces macrophage death by caspase-1-dependent necrosis [J].
Brennan, MA ;
Cookson, BT .
MOLECULAR MICROBIOLOGY, 2000, 38 (01) :31-40
[9]   Acute ethanol administration results in a protective cytokine and neuroinflammatory profile in traumatic brain injury [J].
Chandrasekar, Akila ;
Heuvel, Florian Olde ;
Palmer, Annette ;
Linkus, Birgit ;
Ludolph, Albert C. ;
Boeckers, Tobias M. ;
Relja, Borna ;
Huber-Lang, Markus ;
Roselli, Francesco .
INTERNATIONAL IMMUNOPHARMACOLOGY, 2017, 51 :66-75
[10]   ATP activates a reactive oxygen species-dependent oxidative stress response and secretion of proinflammatory cytokines in macrophages [J].
Cruz, Cristiane M. ;
Rinna, Alessandra ;
Forman, Henry Jay ;
Ventura, Ana L. M. ;
Persechini, Pedro M. ;
Ojcius, David M. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (05) :2871-2879