Differential down-regulation of COX-2 and MMP-13 in human skin fibroblasts by glucosamine-hydrochloride.
BACKGROUND
Evidence suggests anti-inflammatory effects of glucosamine (GS) on inflammatory diseases. COX-2 is an enzyme to produce prostaglandins. MMPs are the family of matrix metalloproteinases degradable of ECM. Excess expression of COX-2 or MMPs involves in skin inflammation.


OBJECTIVE
We evaluated whether GS-HCl modulates expression of COX-2 and/or MMPs by IL-1beta or PMA in human skin fibroblasts (HSF) or keratinocytes (HaCaT).


METHODS
HSF or HaCaT cells were exposed to IL-1beta or PMA without or with GS-HCl. COX-2 or MMPs protein and mRNA expression, respectively, were analyzed by Western blot and RT-PCR. MTS assay was utilized to assess the cytotoxicity of GS-HCl on HSF cells.


RESULTS
In HSF cells, IL-1beta treatment induced COX-2 and MMP-13 expressions in association with activation of ERKs, p38 MAPK, JNKs, and NF-kappaB. PMA treatment also induced COX-2 and MMP-13 expressions in association with p38 MAPK activation. Of interest, treatment with GS-HCl (10mM) led to blockage of p38 MAPK activation, accumulation of 66kDa COX-2 protein variant (without affecting COX-2 mRNA expression), and transcriptional down-regulation of MMP-13 in the IL-1beta- or PMA-treated HSF cells. Distinctly, pharmacological inhibition of p38 MAPK with SB203580 was associated with transcriptional down-regulation of COX-2 and MMP-13 in the IL-1beta- or PMA-treated HSF cells. In addition, the GS-HCl-mediated COX-2 protein modification was observed in both endogenous and PMA-induced COX-2 in HaCaT cells.


CONCLUSIONS
GS-HCl differentially down-regulates COX-2 and MMP-13 expression in the IL-1beta- or PMA-treated human skin fibroblasts via the p38 MAPK-independent COX-2 translational inhibition and the p38 MAPK-dependent MMP-13 transcriptional suppression, respectively.
Hong
Hua
H
Department of Medical Genetic Engineering, Keimyung University School of Medicine, 194 Dongsan-dong, Jung-gu, Daegu, 700-712, South Korea.
Park
Yu-Kyoung
YK
Choi
Mi-Sun
MS
Ryu
Nam-Hee
NH
Song
Dae-Kyu
DK
Suh
Seong-Il
SI
Nam
Ki-Young
KY
Park
Gy-Young
GY
Jang
Byeong-Churl
BC
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002273:Carcinogens / Q000494:pharmacology; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005944:Glucosamine / Q000494:pharmacology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D053583:Interleukin-1beta / Q000494:pharmacology; D015603:Keratinocytes / Q000187:drug effects / Q000201:enzymology; D053509:Matrix Metalloproteinase 13; D061965:Matrix Metalloproteinase Inhibitors; D011494:Protein Kinases / Q000187:drug effects / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D012867:Skin / Q000187:drug effects* / Q000201:enzymology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D002273:Carcinogens; D004791:Enzyme Inhibitors; D007093:Imidazoles; D053583:Interleukin-1beta; D061965:Matrix Metalloproteinase Inhibitors; D011725:Pyridines; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011494:Protein Kinases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D005944:Glucosamine; D013755:Tetradecanoylphorbol Acetate; C093642:SB 203580