Monensin stimulation of arylsulfatase B activity in human chondrocytes.
Secreted and intracellular arylsulfatase B (ASB) activities were measured in normal and osteoarthritic (OA) human chondrocyte cultures in the absence and presence of monensin, ammonium chloride, and chloroquine. Of the three agents added, only monensin produced a significant stimulation of secreted enzyme activity. Osteoarthritic cells consistently exhibited a three-fold higher level of secreted specific ASB activity than did normal cells, with or without monensin. When compared with normal cells, OA cells also consistently exhibited a twofold heightened intracellular specific enzyme activity both in the absence or presence of monensin. With increasing dosage of monensin, secreted and intracellular ASB activity increased for both OA and normal cells. Total enzyme activity of secreted and intracellular ASB was found to be cell density dependent. No inhibition of secreted or intracellular ASB activity was observed for sparsely plated cultures. In contrast to sparse cultures, an inhibition of secreted ASB, with or without monensin, was observed in densely plated cultures. Intracellular total activity was not inhibited by high-density cultures. Secreted ASB activity was found to be time-dependent after passage. Enzyme activity was maximal at 6 h in both OA and normal cells and decreased by the end of 24 h both in serum-free medium and in serum-free medium with monensin. When compared with normal cells, OA cells expressed higher levels of ASB activity under all test conditions. This heightened activity therefore appears to be a property inherent in the OA chondrocyte.
D000368:Aged; D000643:Ammonium Chloride / Q000494:pharmacology; D002358:Cartilage, Articular / Q000166:cytology / Q000201:enzymology*; D002452:Cell Count; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D002803:Chondro-4-Sulfatase / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005663:Furans / Q000494:pharmacology*; D006801:Humans; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology; D008297:Male; D008875:Middle Aged; D008985:Monensin / Q000494:pharmacology*; D010003:Osteoarthritis / Q000201:enzymology; D013429:Sulfatases / Q000378:metabolism*
D005663:Furans; D000643:Ammonium Chloride; D002738:Chloroquine; D008985:Monensin; D013429:Sulfatases; D002803:Chondro-4-Sulfatase