Heat-induced MMP-1 expression is mediated by TRPV1 through PKCalpha signaling in HaCaT cells.
BACKGROUND
Matrix metalloproteinase-1 (MMP-1) is considered a key initiator of collagen degradation in inflammatory responses. A heat-gated channel, transient receptor potential vanilloid type 1 (TRPV1), induces release of proinflammatory mediators. TRPV1 channels have been localized to the epidermis and we have recently suggested that they act as mediators of heat-induced MMP-1. The aim of this study was to investigate the signaling of TRPV1 in MMP-1 regulation by heat shock in human epidermal keratinocytes.


METHODS
Heat shock-induced MMP-1 expression was decreased by treatment with TRPV1 inhibitor. The heat-induced MMP-1 expression was suppressed by Gö6976 [calcium-dependent inhibitor] and staurosporine (ST, broad-spectrum PKC inhibitor), while rottlerin (ROT, calcium-independent PKCdelta inhibitor) had no effect. Also, transfection of PKCalpha siRNA decreased MMP-1 expression, whereas MMP-1 expression was not significantly affected in cells transfected with negative control siRNA, PKCbeta siRNA or PKCdelta siRNA.


RESULTS
We demonstrated that heat shock failed to induce MMP-1 expression in HaCaT cells cultured in calcium-free media. The heat-induced [Ca(2+)](i) increase was inhibited by Gö6976 and ST, but not by ROT. We also found that heat-induced phosphorylation of ERK, JNK and p38 MAPK in HaCaT cells, but capsazepine and ruthenium red had no effect on this activation. In addition to the role of TRPV1 in heat-induced MMP-1 expression, we also found that heat increased TRPV1 proteins in human skin in vivo.


CONCLUSIONS
Our results suggest that TRPV1 mediates heat shock-induced MMP-1 expression via calcium-dependent PKCalpha signaling in HaCaT cells.
Lee
Young Mee
YM
Department of Dermatology, Seoul National University College of Medicine, Clinical Research Institute, Seoul National University Hospital, Seoul National University, Seoul, Korea.
Li
Wen Hai
WH
Kim
Yeon Kyung
YK
Kim
Kyu Han
KH
Chung
Jin Ho
JH
D002118:Calcium / Q000378:metabolism; D002461:Cell Line, Transformed; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D018869:Heat-Shock Response / Q000502:physiology; D006358:Hot Temperature; D006801:Humans; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology*; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D051571:Protein Kinase C-alpha / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D050916:TRPV Cation Channels / Q000378:metabolism*
D050916:TRPV Cation Channels; C494482:TRPV1 protein, human; D051571:Protein Kinase C-alpha; D048049:Extracellular Signal-Regulated MAP Kinases; D020781:Matrix Metalloproteinase 1; D002118:Calcium