title: Lipopolysaccharide/adenosine triphosphate-mediated signal transduction in the regulation of NLRP3 protein expression and caspase-1-mediated interleukin-1β secretion.
abstract: OBJECTIVE
  Reactive oxygen species (ROS) plays a critical role in the regulation of NLRP3 inflammasome activation. However, the ROS-mediated signaling pathways controlling NLRP3 inflammasome activation are not well defined.
  
  
  METHODS
  Using lipopolysaccharide (LPS) and adenosine triphosphate (ATP) activated murine macrophages as the testing model, cytokine release and protein expression were quantified by enzyme-linked immunosorbent assay and Western blot, respectively. ROS was scavenged by N-acetyl cysteine; NADPH oxidase, the major source of ROS, was inhibited by diphenyliodonium, apocynin or gp91-phox siRNA transfection; and protein kinase was inhibited by its specific inhibitor.
  
  
  RESULTS
  LPS-induced NLRP3 protein expression was regulated through the NADPH oxidase/ROS/NF-κB-dependent, JAK2/PI3-kinase/AKT/NF-κB-dependent, and MAPK-dependent pathways, while ATP-induced caspase-1 activation was regulated through the NADPH oxidase/ROS-dependent pathway.
  
  
  CONCLUSIONS
  These results demonstrate that ROS regulates not only the priming stage, but also the activation stage, of NLRP3 inflammasome activation in LPS + ATP-activated macrophages.
authors.affiliation: Department of Biotechnology and Animal Science, National Ilan University, 1 Sec. 1 Shen-Lung Road, Ilan, Taiwan.
authors.forename: Pei-Chun
authors.initials: PC
authors.lastname: Liao
authors.forename: Louis Kuoping
authors.initials: LK
authors.lastname: Chao
authors.forename: Ju-Ching
authors.initials: JC
authors.lastname: Chou
authors.forename: Wei-Chih
authors.initials: WC
authors.lastname: Dong
authors.forename: Chien-Nan
authors.initials: CN
authors.lastname: Lin
authors.forename: Chai-Yi
authors.initials: CY
authors.lastname: Lin
authors.forename: Ann
authors.initials: A
authors.lastname: Chen
authors.forename: Shuk-Man
authors.initials: SM
authors.lastname: Ka
authors.forename: Chen-Lung
authors.initials: CL
authors.lastname: Ho
authors.forename: Kuo-Feng
authors.initials: KF
authors.lastname: Hua
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D020170:Caspase 1 / Q000502:physiology*; D002478:Cells, Cultured; D053583:Interleukin-1beta / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D051379:Mice; D019255:NADPH Oxidases / Q000502:physiology; D016328:NF-kappa B / Q000502:physiology; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D002352:Carrier Proteins; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D017382:Reactive Oxygen Species; D000255:Adenosine Triphosphate; D019255:NADPH Oxidases; D020170:Caspase 1