Flashlamp pulsed-dye laser suppressed TGF-beta1 expression and proliferation in cultured keloid fibroblasts is mediated by MAPK pathway.
OBJECTIVE
Our previous clinical study indicated that transforming growth factor-beta1 (TGF-beta1) and mitogen-activated protein kinases (MAPK) are both involved in keloid regression following flashlamp pulsed-dye laser (PDL). To further characterize of this involvement, this work examined whether PDL suppression of TGF-beta1 expression was mediated through MAPK pathway in cultured keloid fibroblasts (KF).


METHODS
Primary culture of KF harvested from keloid patients received various dosages of PDL treatment in 585-nm wavelength. TGF-beta1 expressions in KF following various dosages of PDL were assessed. Additionally, MAPK pathway activities were studied using the PD98059 (an ERK inhibitor), SB203580 (a p38 kinase inhibitor), and SP600125 (a JNK inhibitor), to determine the role in keloid following PDL treatment. Activator protein-1 (AP-1), a transcription factor of TGF-beta, was analyzed by electrophoretic mobility shift assay (EMSA). Phosphorylated c-Jun, one of the components of AP-1, was also detected.


RESULTS
The observation results demonstrated that optimal dosages of PDL significantly suppressed KF proliferation and TGF-beta1 expression. EMSA study identified PDL downregulation of super-shift of AP-1. Three subtypes of MAPK cascades were augmented between 30 minutes and 4 hours following PDL treatment, particularly phosphorylation of ERK1/2 and p38. Pre-treatment with PD98059, SB203580, but not SP600125, markedly inhibited the downregulating effects of TGF-beta1 and phosphorylated c-Jun expression following PDL treatment.


CONCLUSIONS
PDL induced keloid regression is mediated by triggering MAPK cascades and blockade of AP-1 transcription and TGF-beta expression. Modulation of TGF-beta and MAPK interaction in keloids may provide specific targets for therapeutic intervention.
Kuo
Yur-Ren
YR
Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan. t1207816@ms22.hinet.net
Wu
Wen-Sheng
WS
Wang
Feng-Sheng
FS
D000873:Anthracenes / Q000494:pharmacology; D049109:Cell Proliferation / Q000528:radiation effects*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000528:radiation effects*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases / Q000187:drug effects; D007627:Keloid / Q000473:pathology / Q000532:radiotherapy; D028022:Low-Level Light Therapy; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D010766:Phosphorylation / Q000187:drug effects / Q000528:radiation effects; D011725:Pyridines / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000528:radiation effects; D053773:Transforming Growth Factor beta1 / Q000528:radiation effects*
D000873:Anthracenes; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007093:Imidazoles; D011725:Pyridines; D018808:Transcription Factor AP-1; D053773:Transforming Growth Factor beta1; C432165:pyrazolanthrone; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one