Synoviocytes are more sensitive than cartilage to the effects of minocycline and doxycycline on IL-1alpha and MMP-13-induced catabolic gene responses.
The objective of this study was to determine the primary articular tissue target of doxycycline and minocycline. Synoviocytes-cartilage cocultures (n = 4) were treated with MMP-13 (25 ng/mL medium) or IL-1 (1.0 ng/mL medium) for 24 h. Doxycycline (4.3, 0.43, 0.043 microM) or minocycline (10, 1.0 or 0.1 microM) were then added and cultures were continued for 96 h. Cartilage and media were analyzed for GAG content. Quantitative PCR was used to measure cartilage MMP-3, MMP-13, aggrecan, COL2A1, ADAMTS-4, and ADAMTS-5 expression, and synoviocyte MMP-3, MMP-13, ADAMTS-4, and ADMATS-5 expression. Total and active MMP-3, MMP-13, and ADAMTS 4/5 enzymes were measured in culture medium. All concentrations of doxycycline and minocycline diminished GAG accumulation in the media. All concentrations of minocycline, but only the highest concentration of doxycycline decreased MMP-3 and MMP-13 expression in synoviocytes but not cartilage, and basal ADAMTS-5 mRNA levels in both synoviocytes and cartilage. Only minocycline decreased active MMP-13 protein in synoviocytes. In summary, the protective effects of tetracycline compounds are more pronounced in synoviocytes than cartilage, and following minocycline compared to doxycycline. Studies to determine the molecular mechanism of action of the tetracyclines in synoviocytes might lead to the design of targeted therapeutics for the treatment of OA or RA.
Fortier
Lisa A
LA
Department of Clinical Sciences, Cornell University, VMC C3-181, Ithaca, New York 14853, USA. laf4@cornell.edu
Motta
Tatiana
T
Greenwald
Robert A
RA
Divers
Thomas J
TJ
Mayr
Karen G
KG
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000378:metabolism; D004318:Doxycycline / Q000494:pharmacology*; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006023:Glycoproteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006025:Glycosaminoglycans / Q000032:analysis / Q000378:metabolism; D006736:Horses / Q000502:physiology; D053582:Interleukin-1alpha / Q000494:pharmacology*; D064235:Matrilin Proteins; D053509:Matrix Metalloproteinase 13 / Q000494:pharmacology*; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism; D008911:Minocycline / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D013264:Stifle / Q000166:cytology / Q000187:drug effects; D013583:Synovial Membrane / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D017077:Culture Media, Conditioned; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D006025:Glycosaminoglycans; D053582:Interleukin-1alpha; D064235:Matrilin Proteins; D012333:RNA, Messenger; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3; D008911:Minocycline; D004318:Doxycycline