Department of Biomedicine, Division of Rheumatology, AOUC, Excellence Centre for Research, Transfer and High Education DENOthe, University of Florence, Florence, Italy.
Ann Rheum Dis. 2011 Nov;70(11):2011-21. doi: 10.1136/ard.2011.150607. Epub 2011 Aug 6.
To characterise bone marrow-derived mesenchymal stem cells (MSCs) from patients with systemic sclerosis (SSc) for the expression of factors implicated in MSC recruitment at sites of injury, angiogenesis and fibrosis. The study also analysed whether the production/release of bioactive mediators by MSCs were affected by stimulation with cytokines found upregulated in SSc serum and tissues, and whether MSCs could modulate dermal microvascular endothelial cell (MVEC) angiogenesis.
MSCs obtained from five patients with early severe diffuse SSc (SSc-MSCs) and five healthy donors (H-MSCs) were stimulated with vascular endothelial growth factor (VEGF), transforming growth factor β (TGFβ) or stromal cell-derived factor-1 (SDF-1). Transcript and protein levels of SDF-1 and its receptor CXCR4, VEGF, TGFβ(1) and receptors TβRI and TβRII were evaluated by quantitative real-time PCR, western blotting and confocal microscopy. VEGF, SDF-1 and TGFβ(1) secretion in culture supernatant was measured by ELISA. MVEC capillary morphogenesis was performed on Matrigel with the addition of MSC-conditioned medium.
In SSc-MSCs the basal expression of proangiogenic SDF-1/CXCR4 and VEGF was significantly increased compared with H-MSCs. SSc-MSCs constitutively released higher levels of SDF-1 and VEGF. SDF-1/CXCR4 were upregulated after VEGF stimulation and CXCR4 redistributed from the cytoplasm to the cell surface. VEGF was increased by SDF-1 challenge. VEGF, TGFβ and SDF-1 stimulation upregulated TGFβ(1), TβRI and TβRII in SSc-MSCs. TβRII redistributed from the cytoplasm to focal adhesion contacts. SSc-MSC-conditioned medium showed a greater proangiogenic effect on MVECs than H-MSCs. Experiments with blocking antibodies showed that MSC-derived cytokines were responsible for this potent proangiogenic effect.
SSc-MSCs constitutively overexpress and release bioactive mediators/proangiogenic factors and potentiate dermal MVEC angiogenesis.
描述系统性硬化症（SSc）患者骨髓间充质干细胞（MSCs）的特征，这些特征涉及到损伤部位、血管生成和纤维化中MSC 募集的相关因子。该研究还分析了 MSC 产生/释放生物活性介质是否受到 SSc 血清和组织中上调的细胞因子的刺激影响，以及 MSC 是否可以调节真皮微血管内皮细胞（MVEC）的血管生成。
从 5 例早期严重弥漫性 SSc 患者（SSc-MSCs）和 5 例健康供体（H-MSCs）中获取 MSC，用血管内皮生长因子（VEGF）、转化生长因子β（TGFβ）或基质细胞衍生因子-1（SDF-1）刺激这些 MSC。通过定量实时 PCR、western blot 和共聚焦显微镜评估 SDF-1 及其受体 CXCR4、VEGF、TGFβ(1)和受体 TβRI 和 TβRII 的转录和蛋白水平。通过 ELISA 测量培养上清液中 VEGF、SDF-1 和 TGFβ(1)的分泌。在添加 MSC 条件培养基的情况下，在 Matrigel 上进行 MVEC 毛细血管形态发生。
与 H-MSCs 相比，SSc-MSCs 中促血管生成 SDF-1/CXCR4 和 VEGF 的基础表达显著增加。SSc-MSCs 持续释放更高水平的 SDF-1 和 VEGF。VEGF 刺激后 SDF-1/CXCR4 上调，CXCR4 从细胞质重新分布到细胞膜。SDF-1 刺激增加了 VEGF。VEGF、TGFβ 和 SDF-1 刺激上调了 SSc-MSCs 中的 TGFβ(1)、TβRI 和 TβRII。TβRII 从细胞质重新分布到粘着斑接触点。SSc-MSC 条件培养基对 MVEC 的促血管生成作用强于 H-MSC。用阻断抗体的实验表明，MSC 衍生的细胞因子是这种强烈促血管生成作用的原因。
SSc-MSCs 持续过表达和释放生物活性介质/促血管生成因子，并增强真皮 MVEC 的血管生成。
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