N-acetylcysteine prevents nitric oxide-induced chondrocyte apoptosis and cartilage degeneration in an experimental model of osteoarthritis.
We investigated whether N-acetylcysteine (NAC), a precursor of glutathione, could protect rabbit articular chondrocytes against nitric oxide (NO)-induced apoptosis and could prevent cartilage destruction in an experimental model of osteoarthritis (OA) in rats. Isolated chondrocytes were treated with various concentrations of NAC (0-2 mM). Apoptosis was induced by 0.75 mM sodium nitroprusside (SNP) dehydrate, which produces NO. Cell viability was assessed by MTT assay, while apoptosis was evaluated by Hoechst 33342 and TUNEL staining. Intracellular reactive oxygen species (ROS) and glutathione levels were measured, and expression of p53 and caspase-3 were determined by Western blotting. To determine whether intraarticular injection of NAC prevents cartilage destruction in vivo, cartilage samples of an OA model were subjected to H&E, Safranin O, and TUNEL staining. NAC prevented NO-induced apoptosis, ROS overproduction, p53 up-regulation, and caspase-3 activation. The protective effects of NAC were significantly blocked by buthionine sulfoximine, a glutathione synthetase inhibitor, indicating that the apoptosis-preventing activity of NAC was mediated by glutathione. Using a rat model of experimentally induced OA, we found that NAC also significantly prevented cartilage destruction and chondrocyte apoptosis in vivo. These results indicate that NAC inhibits NO-induced apoptosis of chondrocytes through glutathione in vitro, and inhibits chondrocyte apoptosis and articular cartilage degeneration in vivo.
Nakagawa
Shuji
S
Department of Orthopaedics, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602-8566, Japan.
Arai
Yuji
Y
Mazda
Osam
O
Kishida
Tsunao
T
Takahashi
Kenji A
KA
Sakao
Kei
K
Saito
Masazumi
M
Honjo
Kuniaki
K
Imanishi
Jiro
J
Kubo
Toshikazu
T
D000111:Acetylcysteine / Q000008:administration & dosage*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism; D004195:Disease Models, Animal; D016166:Free Radical Scavengers / Q000008:administration & dosage*; D005978:Glutathione / Q000187:drug effects / Q000378:metabolism; D007270:Injections, Intra-Articular; D008297:Male; D009569:Nitric Oxide / Q000009:adverse effects; D010003:Osteoarthritis / Q000139:chemically induced / Q000503:physiopathology / Q000517:prevention & control*; D011817:Rabbits; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D016159:Tumor Suppressor Protein p53; D009569:Nitric Oxide; D053148:Caspase 3; D005978:Glutathione; D000111:Acetylcysteine