Institute of Anatomy, Faculty of Medicine, National Yang-Ming University, Taipei 112, Taiwan; Department of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei 112, Taiwan.
Stem Cell Laboratory, Department of Medical Research, Taipei Veterans General Hospital, Taipei 112, Taiwan.
J Mol Cell Cardiol. 2014 Jul;72:339-49. doi: 10.1016/j.yjmcc.2014.04.012. Epub 2014 Apr 27.
Mesenchymal stem cells (MSCs) have been shown to secrete cytokines and growth factors required for angiogenesis. Previously, we demonstrated that MSCs expressing HPV16 E6E7 mRNA (E6E7-MSCs) increase life span and differentiation potential and maintain without neoplastic transformation. Whether E6E7-MSCs are sources of molecules for enhancing angiogenesis is unknown. We demonstrated that E6E7-MSC-derived conditioned medium (E6E7-CM) enhanced endothelial cell migration and tube formation compared to primary MSC-derived conditioned medium (primary-CM). Moreover, E6E7-MSCs increased AKT activation and enhanced the release of Interleukin-1β (IL-1β) and vascular endothelial growth factor A (VEGFA). Neutralization of E6E7-CM with antibodies against IL-1β or VEGFA abrogated its effect in enhancing endothelial migration and tube formation. Primary-CM, added with IL-1β and VEGFA, enhanced its ability to increase endothelial migration and tube formation. E6E7-CM was shown to increase the ability to improve blood perfusion in a mouse limb ischemia model. Histological analysis revealed that E6E7-CM prohibited muscle loss or fibrosis and increased endothelial cell counts compared to primary-CM. Similarly, the effects of E6E7-CM in improving perfusion in ischemic limb were also contributed by the increase of IL-1β or VEGFA levels. These results suggest that E6E7-MSCs increase the ability to secrete angiogenic factors via AKT activation, and E6E7-CM is abundant in IL-1β and VEGFA levels and thereby increases the ability to improve blood perfusion and prohibit muscle loss or fibrosis in a mouse limb ischemia model.
间充质干细胞（MSCs）已被证明可分泌血管生成所需的细胞因子和生长因子。此前，我们证明表达 HPV16 E6E7 mRNA 的 MSCs（E6E7-MSCs）可增加寿命和分化潜能，并保持非肿瘤转化。E6E7-MSCs 是否是增强血管生成的分子来源尚不清楚。我们证明 E6E7-MSC 衍生的条件培养基（E6E7-CM）与原代 MSC 衍生的条件培养基（primary-CM）相比，可增强内皮细胞迁移和管状结构形成。此外，E6E7-MSCs 增加 AKT 激活，并增强白细胞介素 1β（IL-1β）和血管内皮生长因子 A（VEGFA）的释放。用针对 IL-1β 或 VEGFA 的抗体中和 E6E7-CM 可消除其增强内皮细胞迁移和管状结构形成的作用。添加 IL-1β 和 VEGFA 的 primary-CM 增强了其增强内皮细胞迁移和管状结构形成的能力。E6E7-CM 被证明可增加改善小鼠肢体缺血模型中血液灌注的能力。组织学分析显示，与 primary-CM 相比，E6E7-CM 可防止肌肉损失或纤维化，并增加内皮细胞计数。同样，E6E7-CM 通过增加 IL-1β 或 VEGFA 水平来改善缺血肢体灌注的作用。这些结果表明，E6E7-MSCs 通过 AKT 激活增加了分泌血管生成因子的能力，E6E7-CM 富含 IL-1β 和 VEGFA 水平，从而增加了改善血液灌注和防止小鼠肢体缺血模型中肌肉损失或纤维化的能力。
