Suppression of cartilage degeneration by intra-articular injection of heparan sulfate 6-O endosulfatase in a mouse osteoarthritis model
Suppression of cartilage degeneration by intra-articular injection of heparan sulfate 6-O endosulfatase in a mouse osteoarthritis model
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DOI：
10.14670/hh-11-838
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发表时间：
2017-07-01
影响因子：
2
通讯作者：
Neo, Masashi
中科院分区：
文献类型：
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作者：
Otsuki, Shuhei;Murakami, Tomohiko;Neo, Masashi
我们以前报道过硫酸乙酰肝素6-O内硫酸酯酶（Sulfs）在关节软骨中表达，并且Sulf-1基因敲除小鼠发生了严重的膝关节骨关节炎。在这项研究中，我们假设关节内注射Sulf-1可以防止软骨退化。在确认1 mg/ml Sulf-1在体外不诱导ATDC 5细胞死亡后，通过定量实时聚合酶链反应测定在Sulf-1（1-100 ng/ml）存在下II型胶原和基质金属蛋白酶（MMP）-13的基因表达。在手术破坏内侧半月板的稳定性后，还将Sulf-1关节内注射到小鼠中以产生骨关节炎模型，并通过番红O和MMP-13染色评价软骨变性。我们还研究了成纤维细胞生长因子2（FGF 2）/细胞外信号调节激酶（Erk）细胞信号通过蛋白质印迹。暴露于Sulf-1在体外增加II型胶原蛋白的表达和降低MMP-13的表达在浓度依赖性的方式。Sulf-1注射到小鼠骨关节炎膝关节显著抑制糖胺聚糖损失和MMP-13表达。Sulf-1和FGF 2显著降低了Erk 1/2信号通路的激活。这些发现表明，Sulf-1通过对FGF 2/Erk 1/2信号传导的影响抑制MMP-13来防止软骨退化。
We previously reported that heparan sulfate 6-O endosulfatases (Sulfs) were expressed in articular cartilage, and that the Sulf-1 knockout mouse developed severe knee osteoarthritis. In this study, we hypothesised that intra-articular injection of Sulf-1 would prevent cartilage degeneration. After confirming that 1 mg/ml Sulf-1 did not induce ATDC5 cell death in vitro, gene expression of type II collagen and matrix metalloproteinase (MMP)-13 in the presence of Sulf-1 (1-100 ng/ml) were determined by quantitative real-time polymerase chain reaction. Sulf-1 was also injected intra-articularly into mice following surgical destabilisation of the medial meniscus to produce a model of osteoarthritis, and cartilage degeneration was evaluated by safranin O and MMP-13 staining. We also investigated fibroblast growth factor 2 (FGF2)/extracellular signal-regulated kinase (Erk) cell signalling by western blotting. Exposure to Sulf-1 in vitro increased type II collagen expression and decreased MMP-13 expression in a concentration-dependent manner. Sulf-1 injection into the mouse osteoarthritic knee significantly suppressed glycosaminoglycan loss and MMP-13 expression. Erk1/2 signalling pathway activation was significantly reduced by Sulf-1 and FGF2. These findings indicate that Sulf-1 prevents cartilage degeneration by suppressing MMP-13 via an effect on FGF2/Erk1/2 signalling.