title: 2-Amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine is an anti-inflammatory TLR-2, -4 and -5 response mediator in human monocytes.
abstract: OBJECTIVE
  To elucidate the influence of 2-amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine (AMBMP), a canonical Wnt/β-catenin pathway activator, on the inflammatory response of TLR-engaged innate cells in vitro.
  Primary human monocytes.
  
  
  METHODS
  AMPMB (0-10 μM), LPS (0-1.0 μg/ml), Pam3CSK4, FSL-1, or S. typhimurium flagellin (0-0.25 μg/ml).
  
  
  METHODS
  TLR-induced cytokine release (TNF, IL-6, IL-12 p40) was monitored by ELISA while Wnt-related signals (GSK3β, p65, IκB, β-catenin) were assessed by Western blot, pharmaceutical inhibition and gene silencing.
  
  
  RESULTS
  AMBMP induced the rapid phosphorylation of NFκB p65 at Ser(536) and abrogated total IκB, accompanied by a subsequent increase in pro-inflammatory cytokine production (TNF, IL-6, IL-12 p40) in otherwise naive monocytes. However, in TLR2, -4 and -5-engaged monocytes, AMBMP-suppressed cytokine production. In the context of LPS stimulation, this occurred concomitant with the phosphorylative inactivation of GSK3β at Ser(9), β-catenin accumulation and abrogation of NFκB p65 phosphorylation. AMBMP-mediated suppression of the TLR4 -induced inflammatory response was reversed by two pharmaceutical Wnt/β-catenin pathway inhibitors, IWP-2 and PNU-74654 and by Wnt3a silencing.
  
  
  CONCLUSIONS
  Herein, we show that AMBMP induces canonical Wnt signaling events and acts as a suppressor of inflammation in surface TLR-engaged primary human monocytes.
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. huizhi.wang@louisville.edu.
authors.forename: Huizhi
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. mark.graves@louisville.edu.
authors.forename: Mark W
authors.initials: MW
authors.lastname: Graves
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. h0zhou03@louisville.edu.
authors.forename: Huaxin
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. zhen.gu@louisville.edu.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Gu
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. rich.lamont@louisville.edu.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Lamont
authors.affiliation: Department of Oral Immunology and Infectious Diseases, University of Louisville, 501 South Preston Street, Louisville, KY, 40292, USA. dascot07@louisville.edu.
authors.forename: David A
authors.initials: DA
authors.lastname: Scott
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D052117:Benzodioxoles / Q000494:pharmacology*; D016207:Cytokines / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000096:biosynthesis / Q000235:genetics; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism*; D061251:Primary Cell Culture; D011743:Pyrimidines / Q000494:pharmacology*; D060490:Receptors, Wnt / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D051195:Toll-Like Receptor 2 / Q000187:drug effects / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000187:drug effects / Q000378:metabolism*; D051198:Toll-Like Receptor 5 / Q000187:drug effects / Q000378:metabolism*
chemical_list: C581356:2-amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine; D000893:Anti-Inflammatory Agents; D052117:Benzodioxoles; D016207:Cytokines; D008070:Lipopolysaccharides; D011743:Pyrimidines; D060490:Receptors, Wnt; C495342:TLR2 protein, human; C495345:TLR4 protein, human; C495346:TLR5 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051198:Toll-Like Receptor 5; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3
keywords: 2-amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl)pyrimidine; Cytokines; GSK3β; Inflammation; TLR; Wnt; β-catenin