Shosaikoto increases calprotectin expression in human oral epithelial cells.
OBJECTIVE
Oral epithelial cells help to prevent against bacterial infection in the oral cavity by producing antimicrobial peptides (AMPs). A broad-spectrum AMP, calprotectin (a complex of S100A8 and S100A9 proteins), is expressed by oral epithelial cells and is up-regulated by interleukin-1alpha (IL-1alpha). Shosaikoto (SST) is a traditional Japanese herbal medicine that has immunomodulatory effects and is reported to enhance the levels of IL-1alpha in epithelial cells. The purpose of this study was to investigate the effect of SST on the expression of calprotectin and other AMPs through the regulation of IL-1alpha in oral epithelial cells.


METHODS
Human oral epithelial cells (TR146) were cultured with SST (at concentrations ranging from 10 to 250 microg/mL) in the presence or absence of anti-IL-1alpha or IL-1 receptor antagonist. The expression of S100A8- and S100A9-specific mRNAs was examined by northern blotting. Calprotectin expression and IL-1alpha secretion were investigated by immunofluorescent staining or ELISA. The expression of other AMPs and IL-1alpha was analyzed by RT-PCR and by quantitative real-time PCR.


RESULTS
Shosaikoto (25 microg/mL) significantly increased the expression of S100A8- and S100A9-specific mRNAs and calprotectin protein. Shosaikoto increased S100A7 expression, but had no effect on the expression of other AMPs. The expression of IL-1alpha-specific mRNA and its protein were slightly increased by SST. A neutralizing antibody against IL-1alpha or IL-1 receptor antagonist inhibited SST up-regulated S100A8/S100A9 mRNA expression.


CONCLUSIONS
These results suggest that SST increases the expression of calprotectin and S100A7 in oral epithelial cells. In response to SST, up-regulation of calprotectin may be partially induced via IL-1alpha.
Hiroshima
Y
Y
Department of Periodontology and Endodontology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan.
Bando
M
M
Kataoka
M
M
Shinohara
Y
Y
Herzberg
M C
MC
Ross
K F
KF
Inagaki
Y
Y
Nagata
T
T
Kido
J
J
D023181:Antimicrobial Cationic Peptides / Q000032:analysis / Q000187:drug effects; D015152:Blotting, Northern; D040501:Calgranulin A / Q000032:analysis / Q000187:drug effects; D040502:Calgranulin B / Q000032:analysis / Q000187:drug effects; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000494:pharmacology*; D004847:Epithelial Cells / Q000187:drug effects; D006801:Humans; D007155:Immunologic Factors / Q000008:administration & dosage / Q000494:pharmacology*; D053582:Interleukin-1alpha / Q000037:antagonists & inhibitors / Q000494:pharmacology; D039841:Leukocyte L1 Antigen Complex / Q000032:analysis / Q000187:drug effects*; D009061:Mouth Mucosa / Q000166:cytology / Q000187:drug effects*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis / Q000187:drug effects; D053573:Receptors, Interleukin-1 Type I / Q000037:antagonists & inhibitors; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects
D023181:Antimicrobial Cationic Peptides; D040501:Calgranulin A; D040502:Calgranulin B; D004365:Drugs, Chinese Herbal; D007155:Immunologic Factors; D053582:Interleukin-1alpha; D039841:Leukocyte L1 Antigen Complex; D012333:RNA, Messenger; D053573:Receptors, Interleukin-1 Type I; C461320:shosaiko-to