PDF(Pubmed)
Abstract：
Understanding the factors which control endothelial cell (EC) function and angiogenesis is crucial for developing the horse as a disease model, but equine ECs remain poorly studied. In this study, we have optimised methods for the isolation and culture of equine aortic endothelial cells (EAoECs) and characterised their angiogenic functions in vitro. Mechanical dissociation, followed by magnetic purification using an anti-VE-cadherin antibody, resulted in EC-enriched cultures suitable for further study. Fibroblast growth factor 2 (FGF2) increased the EAoEC proliferation rate and stimulated scratch wound closure and tube formation by EAoECs on the extracellular matrix. Pharmacological inhibitors of FGF receptor 1 (FGFR1) (SU5402) or mitogen-activated protein kinase (MEK) (PD184352) blocked FGF2-induced extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation and functional responses, suggesting that these are dependent on FGFR1/MEK-ERK signalling. In marked contrast, vascular endothelial growth factor-A (VEGF-A) had no effect on EAoEC proliferation, migration, or tubulogenesis and did not promote ERK1/2 phosphorylation, indicating a lack of sensitivity to this classical pro-angiogenic growth factor. Gene expression analysis showed that unlike human ECs, FGFR1 is expressed by EAoECs at a much higher level than both VEGF receptor (VEGFR)1 and VEGFR2. These results suggest a predominant role for FGF2 versus VEGF-A in controlling the angiogenic functions of equine ECs. Collectively, our novel data provide a sound basis for studying angiogenic processes in horses and lay the foundations for comparative studies of EC biology in horses versus humans.
摘要：
了解控制内皮细胞（EC）功能和血管生成的因素对于将马发展为疾病模型至关重要。但是马ECs的研究仍然很少。在这项研究中,我们优化了分离和培养马主动脉内皮细胞(EAoECs)的方法,并对其体外血管生成功能进行了表征.机械离解,然后使用抗VE-钙粘蛋白抗体进行磁性纯化，导致富含EC的培养物适合进一步研究。成纤维细胞生长因子2（FGF2）增加了EAoEC的增殖率，并刺激了EAoEC在细胞外基质上的划痕伤口闭合和管形成。FGF受体1（FGFR1）（SU5402）或丝裂原活化蛋白激酶（MEK）（PD184352）的药理学抑制剂阻断FGF2诱导的细胞外信号调节激酶1/2（ERK1/2）磷酸化和功能反应，这表明这些依赖于FGFR1/MEK-ERK信号传导。与此形成鲜明对比的是,血管内皮生长因子-A(VEGF-A)对EAoEC增殖无影响,迁移,或肾小管发生，并且不促进ERK1/2磷酸化，表明对这种经典的促血管生成生长因子缺乏敏感性。基因表达分析表明，与人类ECs不同，FGFR1由EAoEC以比VEGF受体(VEGFR)1和VEGFR2高得多的水平表达。这些结果表明FGF2与VEGF-A在控制马ECs的血管生成功能中的主要作用。总的来说，我们的新数据为研究马的血管生成过程提供了良好的基础,并为马与人的EC生物学的比较研究奠定了基础.
