Effects of tetracyclines on the production of matrix metalloproteinases and plasminogen activators as well as of their natural inhibitors, tissue inhibitor of metalloproteinases-1 and plasminogen activator inhibitor-1.
OBJECTIVE
Evaluation of tetracycline effects on the expression of MMP-1, MMP-3, tissue inhibitor(s) of metalloproteinase-1 (TIMP-1), plasminogen activators (PA), and PA inhibitor-1, which are all involved in the ultimate regulation of MMP activity could provide new insight into how tetracyclines achieve their cartilage preserving effects.


METHODS
We used bovine articular chondrocytes cultured in alginate gel beads for our studies which were initially treated with 10 microM tetracyclines in the presence of IL-1. Only significant effects were studied at additional concentrations. Expression of mRNA was analyzed by RT-PCR-ELISA. The activity of enzymes and TIMP was measured by functional assays; whereas, the level of PAI-1 was determined by ELISA.


RESULTS
Treating chondrocytes with IL-1 induced the expression of MMPs and downregulated TIMP-1 but stimulated both the expression of PAs and PAI-1. When tested at 10 microM only minocycline reduced collagenase activity and expression of MMP-1. Further pharmacokinetic analysis revealed IC50 values of 26 microM and 16 microM for the inhibition of collagenase activity and mRNA expression, respectively. Production of MMP-3 was only decreased by tetracycline (IC50 = 45.4 microM). No effects of tetracyclines could be observed on proteoglycan degradation, TIMP activity and the production of PAs, PAI-1, and TIMP-1.


CONCLUSIONS
We conclude that the inhibition of MMPs by tetracyclines occurs mainly via down-regulation of the respective gene expression.
Sadowski
T
T
Institute of Pharmacology and Toxicology, University of Bonn, Germany. thorsten.sadowski@biochem.uni-kiel.de
Steinmeyer
J
J
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism; D005786:Gene Expression Regulation; D007375:Interleukin-1 / Q000494:pharmacology; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis* / Q000235:genetics; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis* / Q000235:genetics; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis*; D010960:Plasminogen Activators / Q000096:biosynthesis*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D013754:Tetracyclines / Q000494:pharmacology*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis* / Q000235:genetics
D007375:Interleukin-1; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D013754:Tetracyclines; D019715:Tissue Inhibitor of Metalloproteinase-1; D010960:Plasminogen Activators; D019278:Matrix Metalloproteinase 3; D020781:Matrix Metalloproteinase 1