成纤维细胞生长因子 23 以 Klotho 非依赖的方式驱动人骨性关节炎软骨细胞中 MMP13 的表达。
Fibroblast Growth Factor 23 drives MMP13 expression in human osteoarthritic chondrocytes in a Klotho-independent manner.
机构信息
UMR 7365 CNRS-Université de Lorraine « Ingénierie Moléculaire et Physiopathologie Articulaire » (IMoPA), Biopôle de l'Université de Lorraine, Campus Biologie-Santé, Vandœuvre-lès-Nancy, France.
UMR 7365 CNRS-Université de Lorraine « Ingénierie Moléculaire et Physiopathologie Articulaire » (IMoPA), Biopôle de l'Université de Lorraine, Campus Biologie-Santé, Vandœuvre-lès-Nancy, France; Département de Chirurgie Orthopédique et Traumatologique, Centre Hospitalier Universitaire, Nancy, France.
出版信息
Osteoarthritis Cartilage. 2016 Nov;24(11):1961-1969. doi: 10.1016/j.joca.2016.06.003. Epub 2016 Jun 13.
OBJECTIVE
Fibroblast Growth Factor 23 (FGF23) may represent an attractive candidate that could participate to the osteoarthritic (OA)-induced phenotype switch of chondrocytes. To address this hypothesis, we investigated the expression of FGF23, its receptors (FGFRs) and co-receptor (Klotho) in human cartilage and studied the effects of rhFGF23 on OA chondrocytes.
METHOD
Gene expression or protein levels were analysed by RT-PCR and immunohistochemistry. Collagenase 3 (MMP13) activity was measured by a fluorescent assay. MAPK signalling pathways were investigated by phosphoprotein array, immunoblotting and the use of selective inhibitors. RNA silencing was performed to confirm the respective contribution of FGFR1 and Klotho.
RESULTS
We showed that the expression of FGF23, FGFR1 and Klotho was up-regulated at both mRNA and protein levels in OA chondrocytes when compared to healthy ones. These overexpressions were markedly elevated in the damaged regions of OA cartilage. When stimulated with rhFGF23, OA chondrocytes displayed an extended expression of FGF23 and of markers of hypertrophy such as MMP13, COL10A1, and VEGF. We demonstrated that FGF23 auto-stimulation was both FGFR1-and Klotho-dependent, whereas the expression of markers of hypertrophy was mainly dependent on FGFR1 alone. Finally, we showed that FGF23-induced MMP13 expression was strongly regulated by the MEK/ERK cascade and to a lesser extent, by the PI-3K/AKT pathway.
CONCLUSION
These results demonstrate that FGF23 sustains differentiation of OA chondrocytes in a Klotho-independent manner.
目的
成纤维细胞生长因子 23（FGF23）可能是一种有吸引力的候选物，它可以参与软骨细胞的骨关节炎（OA）诱导表型转换。为了验证这一假说，我们研究了 FGF23、其受体（FGFRs）和辅助受体（Klotho）在人软骨中的表达，并研究了 rhFGF23 对 OA 软骨细胞的影响。
方法
通过 RT-PCR 和免疫组织化学分析基因表达或蛋白水平。通过荧光测定法测量胶原酶 3（MMP13）活性。通过磷酸化蛋白阵列、免疫印迹和使用选择性抑制剂研究 MAPK 信号通路。通过 RNA 沉默来确认 FGFR1 和 Klotho 的各自贡献。
结果
与健康软骨细胞相比，我们发现 OA 软骨细胞中 FGF23、FGFR1 和 Klotho 的表达在 mRNA 和蛋白水平上均上调。这些过表达在 OA 软骨的损伤区域明显升高。当用 rhFGF23 刺激时，OA 软骨细胞显示出 FGF23 和肥大标志物（如 MMP13、COL10A1 和 VEGF）的延长表达。我们证明 FGF23 自刺激既依赖于 FGFR1 又依赖于 Klotho，而肥大标志物的表达主要仅依赖于 FGFR1。最后，我们表明 FGF23 诱导的 MMP13 表达强烈受 MEK/ERK 级联调控，而在较小程度上受 PI-3K/AKT 途径调控。
结论
这些结果表明，FGF23 以 Klotho 独立的方式维持 OA 软骨细胞的分化。
Zotero 插件