title: Stimulation of EphB2/ephrin-B1 signalling by tumour necrosis factor alpha in human dental pulp stem cells.
abstract: OBJECTIVE
  The aim of this study was to investigate whether in vitro stimulation of dental pulp stem cells (DPSCs) by tumour necrosis factor alpha (TNF-α) would induce secretion of EphB2/ephrin-B1 signalling.
  
  
  METHODS
  Dental pulp stem cells isolated from human dental pulp were treated with TNF-α (5-100 ng/ml) over 2-48 h. EphB2/ephrin-B1 mRNA and protein levels were measured by real-time polymerase chain reaction (RT-PCR) and western blot analysis respectively. Additionally, DPSCs were pre-incubated with TNF-α receptor neutralizing antibodies or infected with nuclear factor-kappa B (NF-ĸB) inhibitor, p38 MAPK inhibitor, Jun N-terminal kinase (JNK) inhibitor and MEK inhibitor before TNF-α treatment. Results were analysed by one-way ANOVA.
  
  
  RESULTS
  Tumour necrosis factor alpha increased EphB2 mRNA expression in DPSCs at concentrations up to 20 ng/ml and ephrin-B1 at concentrations up to 40 ng/ml (P < 0.05). Its mRNA expression reached maximum at 24 h when treated with TNF-α at 20 ng/ml (P < 0.05). EphB2/ephrin-B1 protein expression levels were high at 16 and 24 h as shown by western blotting. Neutralizing antibodies for TNFR1/2 receptors down-regulated EphB2/ephrin-B1 mRNA expression (P < 0.05) and ephrin-B1 protein expression, but not EphB2 protein expression. JNK-inhibitor inhibited EphB2 mRNA expression only (P < 0.05).
  
  
  CONCLUSIONS
  EphB2/ephrin-B1 were invoked in DPSCs with TNF-α treatment via the JNK-dependent pathway, but not NF-ĸB, p38 MAPK or MEK signalling.
authors: Comprehensive Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong, China.
authors: Lifang
authors: L
authors: Zhu
authors: Waruna Lakmal
authors: WL
authors: Dissanayaka
authors: David William
authors: DW
authors: Green
authors: Chengfei
authors: C
authors: Zhang
mesh_terms: D000293:Adolescent; D000328:Adult; D000906:Antibodies / Q000276:immunology; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D036387:Ephrin-B1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D020935:MAP Kinase Signaling System; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D036183:Receptor, EphB2 / Q000235:genetics / Q000378:metabolism*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000494:pharmacology*; D055815:Young Adult; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors
chemical_list: D000906:Antibodies; C484288:EFNB1 protein, human; D036387:Ephrin-B1; D016328:NF-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; C506235:TNFRSF1A protein, human; C415902:TNFRSF1B protein, human; D014409:Tumor Necrosis Factor-alpha; C586310:EPHB2 protein, human; D036183:Receptor, EphB2; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D020929:Mitogen-Activated Protein Kinase Kinases
references: J Periodontol. 2014 Mar;85(3):490-7
references: 23688099
references: Am J Pathol. 2009 Nov;175(5):1905-14
references: 19834065
references: Stem Cells. 2012 Nov;30(11):2472-86
references: 22887987
references: J Endod. 2006 Jan;32(1):39-43
references: 16410066
references: J Dent Res. 2009 Sep;88(9):829-34
references: 19767580
references: Proc Natl Acad Sci U S A. 2011 Jan 25;108(4):1585-90
references: 21209334
references: Cell Prolif. 2011 Oct;44(5):420-7
references: 21951285
references: J Dent. 2010 Sep;38(9):687-97
references: 20580768
references: Cell. 2005 Oct 21;123(2):291-304
references: 16239146
references: Arch Med Res. 2002 Sep-Oct;33(5):482-4
references: 12459320
references: Cell. 2006 Jan 13;124(1):161-73
references: 16413489
references: J Neuroinflammation. 2011 Aug 26;8:106
references: 21867555
references: J Neurosci. 2007 Dec 5;27(49):13481-90
references: 18057206
references: PLoS One. 2013 Sep 09;8(9):e74464
references: 24040255
references: J Biol Chem. 2001 Apr 27;276(17):13771-7
references: 11278471
references: Mol Cell Biol. 2003 Apr;23(8):2871-82
references: 12665585
references: Pharmacol Res. 2008 Aug;58(2):137-47
references: 18602009
references: J Endod. 2013 Oct;39(10):1256-60
references: 24041387
references: Mol Cell Biol. 1990 Nov;10(11):5830-8
references: 2233719
references: Bone. 2011 Mar 1;48(3):533-42
references: 21056708
references: J Dent Res. 2009 Apr;88(4):339-44
references: 19407154
references: J Endod. 2011 Dec;37(12):1663-7
references: 22099901
references: Cytokine Growth Factor Rev. 2003 Jun-Aug;14(3-4):193-209
references: 12787559
references: J Dent Res. 2005 Nov;84(11):994-8
references: 16246929
references: Eur J Cancer. 2006 Feb;42(3):310-8
references: 16406506
references: Stem Cells. 2007 Jan;25(1):156-64
references: 17204606
references: Adv Dent Res. 2011 Jul;23(3):270-4
references: 21677077
references: Blood. 2007 May 1;109(9):4055-63
references: 17197427
references: Cardiovasc Res. 2007 Oct 1;76(1):81-90
references: 17612514