Glucosamine exposure reduces proteoglycan synthesis in primary human endothelial cells in vitro

PURPOSE Glucosamine (GlcN) supplements are promoted for medical reasons, for example, for patients with arthritis and other joint-related diseases. Oral intake of GlcN is followed by uptake in the intestine, transport in the circulation and thereafter delivery to chondrocytes. Here, it is postulated to have an effect on synthesis and turnover of extracellular matrix constituents expressed by these cells. Following uptake in the intestine, serum levels are transiently increased, and the endothelium is exposed to increased levels of GlcN. We investigated the possible effects of GlcN on synthesis of proteoglycans (PGs), an important matrix component, in primary human endothelial cells. METHODS Primary human endothelial cells were cultured in vitro in medium with 5 mM glucose and 0-10 mM GlcN. PGs were recovered and analysed by western blotting, or by SDS-PAGE, gel chromatography or ion-exchange chromatography of (35)S-PGs after (35)S-sulphate labelling of the cells. RESULTS The synthesis and secretion of (35)S-PGs from cultured endothelial cells were reduced in a dose- and time-dependent manner after exposure to GlcN. PGs are substituted with sulphated glycosaminoglycan (GAG) chains, vital for PG function. The reduction in (35)S-PGs was not related to an effect on GAG chain length, number or sulphation, but rather to the total expression of PGs. CONCLUSION Exposure of endothelial cells to GlcN leads to a general decrease in (35)S-PG synthesis. These results suggest that exposure to high levels of GlcN can lead to decreased matrix synthesis, contrary to what has been claimed by supporters of such supplements.