Evidence that human cartilage and chondrocytes do not express calcitonin receptor.
OBJECTIVE
Calcitonin (CT) has been recently shown to exhibit direct protective effects on articular cartilage against joint degenerative disease. It has been proposed that CT might act via the CT receptor (CTR) to activate the cyclic AMP (cAMP) pathway and protect type II collagen degradation. In this study, we investigated the existence of CTR in human articular cartilage and chondrocytes, and examined the potential pharmacological effects and transduction pathway of salmon CT (sCT) in human chondrocytes.


METHODS
Five human articular cartilage samples were examined for the expression of the CTR by polymerase chain reaction (PCR), immunostaining and Western blot analysis. cAMP levels in human chondrocyte stimulated with sCT were assessed by ELISA. The effect of sCT on the gene expression profiles, including aggrecan, type II collagen, MMP-1, MMP-3 and MMP-13, of human chondrocytes was also examined by relative quantitative Real-time PCR.


RESULTS
We failed to detect the CTR at both the transcriptional and protein levels in human chondrocytes and cartilage tissue by PCR, immunostaining and Western blotting. cAMP levels were significantly elevated in human chondrocytes by forskolin (100muM) to more than 10-fold (P<0.001), however, were not induced by sCT (10(-7)M, 10(-8)M, 10(-9)M). Real-time PCR analysis demonstrated that sCT slightly reduced the gene expression of MMPs, although this effect was not statistically significant.


CONCLUSIONS
In contrary to previous reports, our data indicate that human cartilage and chondrocytes do not express CTR. Furthermore, sCT does not appear to have direct effects on human chondrocytes. We propose that the chondroprotective effect of CT observed in vivo may be indirect via its impact on subchondral bone resorptive activity of osteoclasts.
Lin
Z
Z
Centre for Orthopaedic Research, School of Surgery and Pathology, University of Western Australia, Western Australia, Australia.
Pavlos
N J
NJ
Cake
M A
MA
Wood
D J
DJ
Xu
J
J
Zheng
M H
MH
D000818:Animals; D050071:Bone Density Conservation Agents / Q000494:pharmacology*; D002116:Calcitonin / Q000378:metabolism / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007118:Immunoassay; D066298:In Vitro Techniques; D018003:Receptors, Calcitonin / Q000032:analysis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012474:Salmon
D050071:Bone Density Conservation Agents; D018003:Receptors, Calcitonin; D002116:Calcitonin; D000242:Cyclic AMP