The influence of Celecoxib on matrix synthesis by chondrocytes under mechanical stress in vitro.
To assess effects of Celecoxib, selective cyclo-oxygenase (COX)-2 inhibitor, on matrix synthesis by chondrocytes under mechanical stress in vitro. Chondrocytes from 7-day-old rat articular cartilage were cultured. Cyclic mechanical stress (0.5 Hz, 7% elongation) was loaded using a Flexercell strain unit in the presence of Celecoxib, Indomethacin, and Ketoprofen. Concentrations of chondroitin sulfate (CS) in culture media were measured by high-performance liquid chromatography. Nitric oxide (NO) formation was detected by measuring NO2-accumulation in culture supernatants during the Griess reaction, using sodium nitrite as a standard. Levels of mRNA for aggrecan (AGC), type II collagen (CII), COX-2, and membrane-associated prostaglandin E synthase-1 (mPGES-1) were measured using real time reverse transcriptase-polymerase chain reaction. Under mechanical stress, levels of AGC and CII mRNA were decreased, while COX-2 mRNA levels were increased. Of the three drugs tested, only Celecoxib prevented the decrease of AGC mRNA and inhibited the release of CS. In addition, Celecoxib suppresses the level of mPGES-1 mRNA. These findings show that excessive mechanical stress decreases synthesis of matrix components in chondrocytes through a NO-regulated pathway, and suggest that Celecoxib has a protective effect on matrix metabolism.
Iimoto
Seiji
S
Department of Orthopedic Surgery, Ehime University School of Medicine, Toon City, Ehime, Japan. mesy@m.ehime-u.ac.jp
Watanabe
Shohei
S
Takahashi
Toshiaki
T
Shimizu
Akira
A
Yamamoto
Haruyasu
H
D053674:Aggrecans; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology; D001857:Bone Matrix / Q000187:drug effects* / Q000378:metabolism*; D000068579:Celecoxib; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D019746:Intramolecular Oxidoreductases / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D000071876:Prostaglandin-E Synthases; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics; D011509:Proteoglycans / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013314:Stress, Mechanical; D013449:Sulfonamides / Q000494:pharmacology*
C508321:Acan protein, rat; D053674:Aggrecans; D000894:Anti-Inflammatory Agents, Non-Steroidal; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D011720:Pyrazoles; D012333:RNA, Messenger; D013449:Sulfonamides; D009569:Nitric Oxide; D002809:Chondroitin Sulfates; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D019746:Intramolecular Oxidoreductases; D000071876:Prostaglandin-E Synthases; C497818:Ptges protein, rat; D000068579:Celecoxib