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Hypoxia and hypoxia-related genes are important factors in articular chondrocytes during cartilage homeostasis and osteoarthritis. We have investigated the various apoptotic factors that show significance in synovial fluid obtained from normal and experimental osteoarthritic animal models and have evaluated the effect of hypoxia on articular chondrocyte apoptosis induced by these apoptotic factors. Mature beagle dogs underwent surgical transections of ligaments and medial meniscectomies to explore the underlying mechanisms of osteoarthritis. Cartilage and synovial fluid obtained from normal animals and those with osteoarthritis were evaluated via proteasome inhibition, tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) protein expression, mitochondrial transmembrane potential and levels of reactive oxygen species. Canine chondrocytes were exposed to the proteasome inhibitor N-acetyl-Leu-Leu-Norleu-al and treated with recombinant TRAIL protein under normoxic and hypoxic conditions, measuring chondrocyte cell viability, proteasome activity and levels of apoptotic factors. TRAIL protein expression and ubiquitinated proteins were increased significantly, but the proteasome activity in the synovial fluid of osteoarthritic joints relative to that in normal joints was not. Primary cultured articular chondrocytes cotreated with the proteasome inhibitor and TRAIL progressed to severe apoptosis under normoxic conditions, but the sensitization caused by the combined treatment was suppressed by exposure to hypoxia. Caspase-8 activation, c-Jun N-terminal kinase phosphorylation, the mitochondrial transmembrane potential and the generation of reactive oxygen species involved in cell death regulation were significantly inhibited under hypoxic conditions. These findings suggest that proteasome inhibition and TRAIL may be possible mechanisms in cartilage degradation and joint-related diseases. Furthermore, the maintenance of hypoxic conditions or therapy with hypoxia-related genes in the joint may be successful for the treatment of joint-related diseases, including osteoarthritis.