title: Keratinocytes express cytokines and nerve growth factor in response to neuropeptide activation of the ERK1/2 and JNK MAPK transcription pathways.
abstract: Sensory neurons innervating the skin can release neuropeptides that are believed to modulate cellular proliferation, wound healing, pigmentation, and keratinocyte innate immune responses. While the ability of neuropeptides to stimulate keratinocyte production of inflammatory mediators has been demonstrated, there is no information concerning the mechanisms by which neuropeptide activation of keratinocyte cell surface receptors ultimately leads to the up-regulation of mediator production. In this study we used a keratinocyte cell line to identify the presence of substance P (SP) and calcitonin gene-related peptide (CGRP) receptors on keratinocytes and examined the effects of SP and CGRP stimulation on keratinocyte neuropeptide signaling, cell proliferation, and interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), and nerve growth factor (NGF) expression. Neuropeptide stimulation caused an up-regulation of neuropeptide receptor expression in keratinocytes and a dramatic increase in keratinocyte secretion of SP and CGRP, suggesting possible autocrine or paracrine stimulatory effects and amplification of neuropeptide signaling. Both SP and CGRP concentration-dependently stimulated cellular proliferation and the expression and secretion of inflammatory cytokines and NGF in keratinocytes. SP also activated all 3 families of mitogen activated protein kinase (MAPK) and nuclear factor κB (NFκB) in keratinocytes, while CGRP only activated p38 and extracellular signal related kinase1/2 (ERK1/2) MAPKs. Neuropeptide stimulated inflammatory mediatory production in keratinocytes was reversed by ERK1/2 and JNK inhibitors. The current study is the first to observe; 1) that CGRP stimulates keratinocyte expression of CGRP and its receptor complex, 2) that SP and CGRP stimulate IL-6 and TNF-α secretion in keratinocytes, 3) that SP activated all three MAPK families and the NFκB transcriptional signaling pathway in keratinocytes, and 4) that SP and CGRP stimulated inflammatory mediator production in keratinocytes is dependent on ERK1/2 and JNK activation. These studies provide evidence suggesting that disruption of ERK1/2 and JNK signaling may potentially be an effective therapy for inflammatory skin diseases and pain syndromes mediated by exaggerated sensory neuron-keratinocyte signaling.
authors: Physical Medicine and Rehabilitation Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA.|Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA.|Department of Anesthesiolgy, Stanford University School of Medicine, Stanford, CA.
authors: Xiaoyou
authors: X
authors: Shi
authors: Physical Medicine and Rehabilitation Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA.
authors: Liping
authors: L
authors: Wang
authors: Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA.|Department of Anesthesiolgy, Stanford University School of Medicine, Stanford, CA.
authors: J David
authors: JD
authors: Clark
authors: Physical Medicine and Rehabilitation Service, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA.
authors: Wade S
authors: WS
authors: Kingery
mesh_terms: D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D007211:Indoles / Q000494:pharmacology; D018836:Inflammation Mediators / Q000378:metabolism; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism*; D064729:Neurokinin-1 Receptor Antagonists / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D051381:Rats; D058262:Receptor Activity-Modifying Protein 1 / Q000235:genetics / Q000378:metabolism; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000235:genetics / Q000378:metabolism; D013373:Substance P / Q000494:pharmacology / Q000502:physiology; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: C096759:1-(N-(2-methoxybenzyl)acetylamino)-3-(1H-indol-3-yl)-2-(N-(2-(4-(piperidin-1-yl)piperidin-1-yl)acetyl)amino)propane; D016207:Cytokines; D007211:Indoles; D018836:Inflammation Mediators; D009414:Nerve Growth Factors; D064729:Neurokinin-1 Receptor Antagonists; D010880:Piperidines; C545508:Ramp1 protein, rat; D058262:Receptor Activity-Modifying Protein 1; D018015:Receptors, Calcitonin Gene-Related Peptide; D013373:Substance P; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D015740:Calcitonin Gene-Related Peptide
keywords: Calcitonin gene-related peptide; Cytokines; Keratinocyte; Mitogen activated protein kinases; Nerve growth factor; Substance P
references: Physiol Rev. 2006 Oct;86(4):1309-79
references: 17015491
references: Mol Pain. 2007 Oct 02;3:28
references: 17908329
references: J Invest Dermatol. 1993 Aug;101(2):118-26
references: 7688400
references: Biol Reprod. 2002 Aug;67(2):668-73
references: 12135912
references: Pain. 2010 Dec;151(3):843-852
references: 20934254
references: J Invest Dermatol. 1988 Mar;90(3):399-401
references: 2450147
references: J Pain. 2002 Feb;3(1):3-11
references: 14622848
references: J Pharmacol Exp Ther. 1997 Feb;280(2):774-85
references: 9023291
references: Am J Physiol Cell Physiol. 2008 Jun;294(6):C1586-96
references: 18434625
references: Pain. 2008 Aug 15;138(1):47-60
references: 18083307
references: Am J Respir Cell Mol Biol. 1997 Jul;17(1):3-9
references: 9224203
references: Cell Death Differ. 2000 Jun;7(6):566-73
references: 10822280
references: Ann Rheum Dis. 2008 Jul;67(7):909-16
references: 17827184
references: Pain. 2001 Apr;91(3):251-257
references: 11275381
references: J Invest Dermatol. 2001 Nov;117(5):1075-82
references: 11710915
references: Exp Neurol. 2003 Sep;183(1):197-204
references: 12957502
references: Cell Biol Int. 2009 Nov;33(11):1144-8
references: 19651223
references: Exp Dermatol. 2010 Feb;19(2):117-22
references: 19889021
references: Biochim Biophys Acta. 2003 Dec 7;1643(1-3):75-83
references: 14654230
references: Pain. 2011 Sep;152(9):2036-2051
references: 21641113
references: Arch Dermatol Res. 2010 Jan;302(1):5-17
references: 19756672
references: J Invest Dermatol. 1993 Nov;101(5):646-51
references: 8228323
references: Exp Cell Res. 2001 Nov 1;270(2):199-213
references: 11640884
references: Br J Dermatol. 2006 Oct;155(4):657-62
references: 16965412
references: Bone. 2009 Aug;45(2):309-20
references: 19379851
references: J Immunol. 2000 Nov 15;165(10):5606-11
references: 11067916
references: Pain. 2009 Aug;144(3):278-286
references: 19464118
references: J Invest Dermatol. 1983 Feb;80(2):124-30
references: 6185597
references: Methods Mol Biol. 2005;289:273-86
references: 15502192
references: Cytokine. 2002 Apr 21;18(2):72-80
references: 12096921
references: Eur J Pharmacol. 2000 Jun 16;398(2):309-15
references: 10854844
references: Neuropeptides. 2006 Aug;40(4):251-63
references: 16904178
references: Connect Tissue Res. 2005;46(3):153-8
references: 16147857
references: Nature. 1994 Dec 22-29;372(6508):739-46
references: 7997261
references: Expert Opin Biol Ther. 2010 Oct;10(10):1427-39
references: 20738210
references: J Neuroinflammation. 2012 Jul 23;9:181
references: 22824437
references: Biochem Biophys Res Commun. 1999 Sep 24;263(2):327-33
references: 10491292
references: J Invest Dermatol. 2013 Jun;133(6):1620-8
references: 23283070
references: Mol Pain. 2012 Nov 28;8:85
references: 23191958
references: J Pharmacol Exp Ther. 1995 Nov;275(2):737-44
references: 7473161
references: Lancet Neurol. 2011 Jul;10(7):637-48
references: 21683929
references: Anesth Analg. 2011 Jul;113(1):175-83
references: 21596883