OXIDATION OF ARTICULAR-CARTILAGE GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE (G3PDH) OCCURS INVIVO DURING CARRAGEENAN-INDUCED ARTHRITIS

Articular cartilage proteoglycan biosynthesis was substantially inhibited by the competitive glycolytic inhibitor 2-deoxyglucose (approximately 65% at 100 mM), but to a much lesser degree (approximately 10%) by the oxidative phosphorylation uncoupler, 2,4-dinitrophenol. These results confirm that articular cartilage proteoglycan synthesis mostly utilises ATP which is generated by glycolysis. In addition, we have utilised the loss of the relatively specific labelling of glyceraldehyde-3-phosphate dehydrogenase (G3PDH) by [H-3]-iodoacetic acid to show that rabbit articular G3PDH is oxidised in vivo during the animal model of acute arthritis, carrageenin-induced arthritis, in the same way as we have previously shown that cartilage G3PDH is oxidised after in vitro exposure to sublethal doses of H2O2. The oxidation of rabbit G3PDH in vivo (18 hr post-injection) corresponds with the maximal influx of PMNL cells into the arthritic synovial fluid [1] and with substantial inhibition of proteoglycan core protein synthesis [2,3]. We propose that H2O2 released from "activated" PMNLs and macrophages is responsible for the "down-regulation" of biosynthetic processes found in cartilage during acute inflammation.