Histamine induces human beta-defensin-3 production in human keratinocytes.
BACKGROUND
The antimicrobial peptide human beta-defensin-3 (hBD-3) is produced by epidermal keratinocytes, and promotes cutaneous antimicrobial defense, inflammation, and wound repair. hBD-3 induces histamine release from mast cells. We previously showed that histamine enhanced transcriptional activity of activator protein-1 (AP-1) in human keratinocytes by inducing the expression of AP-1 component c-Fos via the activation of extracellular signal-regulated kinase (ERK) through H1 receptors.


OBJECTIVE
To examine in vitro effects of histamine on hBD-3 production in normal human keratinocytes.


METHODS
The hBD-3 production was examined by enzyme-linked immunosorbent assays and reverse transcription-polymerase chain reaction. The transcriptional activities were analyzed by dual luciferase assays. The phosphorylation of proteins was examined by Western blotting.


RESULTS
Histamine enhanced hBD-3 secretion and mRNA expression in keratinocytes. The histamine-induced hBD-3 production was suppressed by H1 antagonist pyrilamine and antisense oligonucleotides against signal transducer and activator of transcription 3 (STAT3) and AP-1 components c-Jun and c-Fos. Histamine enhanced STAT3 transcriptional activity and induced tyrosine and serine phosphorylation of STAT3. The former was suppressed by Janus kinase 2 (JAK2) inhibitor AG490, while the latter was suppressed by mitogen-activated protein kinase kinase (MEK) inhibitor PD98059; both were suppressed by pyrilamine. AG490 and PD98059 suppressed histamine-induced hBD-3 production and STAT3 activity. Histamine induced tyrosine phosphorylation of JAK2, and pyrilamine suppressed the phosphorylation.


CONCLUSIONS
It is suggested that histamine induces hBD-3 production in human keratinocytes through H1 receptors by activating STAT3 and AP-1 via JAK2 and MEK/ERK. Histamine may promote cutaneous antimicrobial defense, inflammation, and wound repair through hBD-3.
Ishikawa
Takeko
T
Department of Dermatology, Teikyo University School of Medicine, 2-11-1, Kaga, Itabashi-Ku, Tokyo 173-8605, Japan. takeko@med.teikyo-u.ac.jp
Kanda
Naoko
N
Hau
Carren S
CS
Tada
Yayoi
Y
Watanabe
Shinichi
S
D015153:Blotting, Western; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005419:Flavonoids / Q000494:pharmacology; D006632:Histamine / Q000378:metabolism*; D006634:Histamine H1 Antagonists / Q000494:pharmacology; D006801:Humans; D007231:Infant, Newborn; D053614:Janus Kinase 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D020930:MAP Kinase Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D008297:Male; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011738:Pyrilamine / Q000494:pharmacology; D034622:RNA Interference; D011969:Receptors, Histamine H1 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D014443:Tyrosine; D020032:Tyrphostins / Q000494:pharmacology; D015854:Up-Regulation; D023083:beta-Defensins / Q000235:genetics / Q000378:metabolism*
C422950:DEFB103A protein, human; D005419:Flavonoids; D006634:Histamine H1 Antagonists; D047428:Protein Kinase Inhibitors; D011969:Receptors, Histamine H1; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D018808:Transcription Factor AP-1; D020032:Tyrphostins; C095512:alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide; D023083:beta-Defensins; D014443:Tyrosine; D006632:Histamine; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D020930:MAP Kinase Kinase Kinases; D011738:Pyrilamine; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one