Inhibition of interleukin-1beta-induced matrix metalloproteinases 1 and 13 production in human osteoarthritic chondrocytes by prostaglandin D2.
OBJECTIVE
To investigate the effects of prostaglandin D2 (PGD2) on interleukin-1beta (IL-1beta)-induced matrix metalloproteinase 1 (MMP-1) and MMP-13 expression in human chondrocytes and the signaling pathways involved in these effects.


METHODS
Chondrocytes were stimulated with IL-1 in the presence or absence of PGD2, and expression of MMP-1 and MMP-13 proteins was evaluated by enzyme-linked immunosorbent assay. Messenger RNA (mRNA) expression and promoter activity were analyzed by real-time reverse transcription-polymerase chain reaction and transient transfections, respectively. The role of the PGD2 receptors D prostanoid receptor 1 (DP1) and chemoattractant receptor-like molecule expressed on Th2 cells (CRTH2) was evaluated using specific agonists and antibody-blocking experiments. The contribution of the cAMP/protein kinase A (PKA) pathway was determined using cAMP-elevating agents and PKA inhibitors.


RESULTS
PGD2 decreased in a dose-dependent manner IL-1-induced MMP-1 and MMP-13 protein and mRNA expression as well as their promoter activation. DP1 and CRTH2 were expressed and functional in chondrocytes. The effect of PGD2 was mimicked by BW245C, a selective agonist of DP1, but not by 13,14-dihydro-15-keto-PGD2, a selective agonist of CRTH2. Furthermore, treatment with an anti-DP1 antibody reversed the effect of PGD2, indicating that the inhibitory effect of PGD2 is mediated by DP1. The cAMP-elevating agents 8-Br-cAMP and forskolin suppressed IL-1-induced MMP-1 and MMP-13 expression, and the PKA inhibitors KT5720 and H89 reversed the inhibitory effect of PGD2, suggesting that the effect of PGD2 is mediated by the cAMP/PKA pathway.


CONCLUSIONS
PGD2 inhibits IL-1-induced production of MMP-1 and MMP-13 by chondrocytes through the DP1/cAMP/PKA signaling pathway. These data also suggest that modulation of PGD2 levels in the joint may have therapeutic potential in the prevention of cartilage degradation.
Zayed
Nadia
N
Centre Hospitalier de l'Université de Montréal, Notre Dame Hospital, and University of Montreal, Montreal, Quebec, Canada.
Afif
Hassan
H
Chabane
Nadir
N
Mfuna-Endam
Leandra
L
Benderdour
Mohamed
M
Martel-Pelletier
Johanne
J
Pelletier
Jean-Pierre
JP
Motiani
Rajender K
RK
Trebak
Mohamed
M
Duval
Nicolas
N
Fahmi
Hassan
H
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000368:Aged; D000906:Antibodies / Q000494:pharmacology; D002227:Carbazoles; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000201:enzymology*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis*; D053509:Matrix Metalloproteinase 13 / Q000096:biosynthesis*; D010003:Osteoarthritis / Q000201:enzymology*; D015230:Prostaglandin D2 / Q000031:analogs & derivatives / Q000494:pharmacology*; D011758:Pyrroles; D012333:RNA, Messenger / Q000032:analysis; D011971:Receptors, Immunologic / Q000502:physiology; D018102:Receptors, Leukotriene B4 / Q000819:agonists / Q000032:analysis; D011982:Receptors, Prostaglandin / Q000819:agonists / Q000276:immunology / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013449:Sulfonamides / Q000494:pharmacology; D018418:Th2 Cells / Q000737:chemistry
D000906:Antibodies; D002227:Carbazoles; D053583:Interleukin-1beta; D007546:Isoquinolines; C513240:LTB4R protein, human; D011758:Pyrroles; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D018102:Receptors, Leukotriene B4; D011982:Receptors, Prostaglandin; D013449:Sulfonamides; C501562:prostanoid D receptor 1, human; D005576:Colforsin; D015124:8-Bromo Cyclic Adenosine Monophosphate; C057416:KT 5720; C034832:13,14-dihydro-15-ketoprostaglandin D2; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D053509:Matrix Metalloproteinase 13; D020781:Matrix Metalloproteinase 1; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D015230:Prostaglandin D2; C036293:prostaglandin D2 receptor