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首页> 外文期刊>Osteoarthritis and cartilage >Evidence that human cartilage and chondrocytes do not express calcitonin receptor.
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Evidence that human cartilage and chondrocytes do not express calcitonin receptor.

机译：人类软骨和软骨细胞不表达降钙素受体的证据。

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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. CONCLUSION: 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.
机译：目的：降钙素（CT）最近被证明对关节软骨具有抗关节退行性疾病的直接保护作用。已经提出CT可能通过CT受体（CTR）起作用以激活环状AMP（cAMP）途径并保护II型胶原蛋白降解。在这项研究中，我们调查了人类关节软骨和软骨细胞中CTR的存在，并研究了鲑鱼CT（sCT）在人类软骨细胞中的潜在药理作用和转导途径。方法：通过聚合酶链反应（PCR），免疫染色和Western blot分析检测五种人关节软骨样品中CTR的表达。通过ELISA评估了用sCT刺激的人软骨细胞中的cAMP水平。还通过相对定量实时PCR检查了sCT对基因表达谱的影响，包括聚集蛋白聚糖，II型胶原，MMP-1，MMP-3和MMP-13。结果：我们无法通过PCR，免疫染色和Western印迹检测到人类软骨细胞和软骨组织中转录水平和蛋白质水平的CTR。 cAMP水平在人软骨细胞中被福司可林（100μM）显着升高至10倍以上（P <0.001），但是，不是由sCT诱导的（10（-7）M，10（-8）M，10（- 9）M）。实时PCR分析表明，sCT稍微降低了MMPs的基因表达，尽管这种作用在统计学上并不显着。结论：与以前的报道相反，我们的数据表明人软骨和软骨细胞不表达CTR。此外，sCT似乎对人类软骨细胞没有直接影响。我们建议在体内观察到的CT的软骨保护作用可能是通过其对破骨细胞软骨下骨吸收活性的影响而间接产生的。

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