Mechanistic study of the proangiogenic effect of osteoprotegerin
Mechanistic study of the proangiogenic effect of osteoprotegerin
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DOI：
10.1007/s10456-013-9337-x
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发表时间：
2013-07-01
期刊：
影响因子：
9.8
通讯作者：
Boisson-Vidal, Catherine
中科院分区：
文献类型：
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作者：
Benslimane-Ahmim, Zahia;Poirier, Florence;Boisson-Vidal, Catherine
骨保护素 (OPG) 是一种可溶性肿瘤坏死因子受体超家族成员，可抑制 RANKL 介导的破骨细胞生成。我们之前报道过 OPG 在体外增强内皮集落形成细胞 (ECFC) 的促血管生成特性，并在体内促进血管生成。在这里，我们研究了 OPG 如何促进新生血管形成。蛋白质组学实验表明，OPG预处理以两种方式影响ECFCs蛋白表达，其中23个点下调，6个点上调。这些点对应于参与细胞运动、粘附、信号转导和细胞凋亡的蛋白质。与这些蛋白质组学结果一致，我们发现 OPG 在剪切应力条件下诱导 ECFC 粘附到活化的内皮上，促进纤连蛋白和胶原的中间粘附，但不促进焦点粘附。 OPG 处理诱导 ECFC 细胞骨架重组，出现迁移表型特征的细胞突起。这些效应与 FAK 磷酸化降低和整合素 α(V)β(3) 表达增强相关。治疗 48 小时后，OPG 显着降低 caspase-3/7 活性并维持 ECFC 活力。 ECFC 与 CXCR4 拮抗剂 AMD-3100 一起孵育以及事先破坏硫酸乙酰肝素蛋白聚糖可显着减弱所有这些效应。 OPG 的促血管生成特性似乎是由蛋白聚糖 syndecan-1 介导的，尽管缺乏肝素结合结构域的 OPG 1-194 在体外和体内仍具有促血管生成作用。这些结果表明OPG可能通过与HSPGs/syndecan-1结合而与ECFCs相互作用，从而诱导抗粘附作用并通过SDF-1/CXCR4依赖性途径促进ECFCs迁移。
Osteoprotegerin (OPG), a soluble tumour necrosis factor receptor superfamily member, inhibits RANKL-mediated osteoclastogenesis. We have previously reported that OPG enhances the proangiogenic properties of endothelial colony-forming cells (ECFCs) in vitro, and promotes vasculogenesis in vivo. Here we investigated how OPG promotes neovascularisation. Proteomic experiments showed that OPG pretreatment affected ECFCs protein expression in two ways, 23 spots being down-regulated and 6 upregulated. These spots corresponded to proteins involved in cell motility, adhesion, signal transduction and apoptosis. In keeping with these proteomic results, we found that OPG induced ECFCs adhesion to activated endothelium in shear stress conditions, promoting intermediate but not focal adhesion to fibronectin and collagen. Treatment with OPG induced a reorganization of the ECFCs cytoskeleton, with the emergence of cell protrusions characteristic of a migratory phenotype. These effects correlated with decreased FAK phosphorylation and enhanced integrin alpha(V)beta(3) expression. OPG drastically reduced caspase-3/7 activities and maintained ECFCs viability after 48 h of treatment. All these effects were significantly attenuated by ECFCs incubation with the CXCR4 antagonist AMD-3100, and by prior heparan sulphate proteoglycan disruption. The proangiogenic properties of OPG appeared to be mediated by the proteoglycan syndecan-1, although OPG 1-194 lacking its heparin-binding domain still had pro-vasculogenic effects in vitro and in vivo. These results suggest that OPG may interact with ECFCs by binding to HSPGs/syndecan-1, thereby induce an anti-adhesive effect and promoting ECFCs migration through a SDF-1/CXCR4 dependent pathway.