Neuromuscular Immunopathology Research Laboratory, Department of Neurology, Baylor College of Medicine, One Baylor Plaza, Mailstop NB302, Houston, TX 77030-3411, USA.
Cell Mol Neurobiol. 2013 Aug;33(6):789-801. doi: 10.1007/s10571-013-9946-3. Epub 2013 May 26.
Several mitogens such as vascular endothelial growth factor (VEGF) have been implicated in mammalian vascular proliferation and repair. However, the molecular mediators of human blood-nerve barrier (BNB) development and specialization are unknown. Primary human endoneurial endothelial cells (pHEndECs) were expanded in vitro and specific mitogen receptors detected by western blot. pHEndECs were cultured with basal medium containing different mitogen concentrations with or without heparin. Non-radioactive cell proliferation, Matrigel(™)-induced angiogenesis and sterile micropipette injury wound healing assays were performed. Proliferation rates, number and total length of induced microvessels, and rate of endothelial cell monolayer wound healing were determined and compared to basal conditions. VEGF-A165 in the presence of heparin, was the most potent inducer of pHEndEC proliferation, angiogenesis, and wound healing in vitro. 1.31 nM VEGF-A165 induced ~110 % increase in cell proliferation relative to basal conditions (∼51 % without heparin). 2.62 pM VEGF-A165 induced a three-fold increase in mean number of microvessels and 3.9-fold increase in total capillary length/field relative to basal conditions. In addition, 0.26 nM VEGF-A165 induced ∼1.3-fold increased average rate of endothelial wound healing 4-18 h after endothelial monolayer injury, mediated by increased cell migration. VEGF-A165 was the only mitogen capable of complete wound closure, occurring within 30 h following injury via increased cell proliferation. This study demonstrates that VEGF-A165, in the presence of heparin, is a potent inducer of pHEndEC proliferation, angiogenesis, and wound healing in vitro. VEGF-A165 may be an important mitogen necessary for human BNB development and recovery in response to peripheral nerve injury.
几种有丝分裂原，如血管内皮生长因子（VEGF），已被牵连到哺乳动物血管增殖和修复中。然而，人血神经屏障（BNB）发育和特化的分子介质尚不清楚。原代人末梢神经内皮细胞（pHEndEC）在体外扩增，并通过 Western blot 检测到特定的有丝分裂原受体。pHEndEC 用含不同有丝分裂原浓度的基础培养基培养，有或无肝素。进行非放射性细胞增殖、Matrigel（™）诱导的血管生成和无菌微管损伤伤口愈合测定。测定并比较增殖率、诱导的微血管数量和总长度以及内皮细胞单层伤口愈合率。在肝素存在的情况下，VEGF-A165 是体外诱导 pHEndEC 增殖、血管生成和伤口愈合的最有效诱导剂。与基础条件相比，1.31 nM VEGF-A165 诱导细胞增殖增加约 110%（无肝素时约 51%）。2.62 pM VEGF-A165 诱导平均微血管数量增加三倍，总毛细血管长度/视野增加 3.9 倍，与基础条件相比。此外，0.26 nM VEGF-A165 诱导内皮单层损伤后 4-18 小时内皮伤口愈合平均速率增加约 1.3 倍，这是通过增加细胞迁移介导的。VEGF-A165 是唯一能够完全闭合伤口的有丝分裂原，在损伤后 30 小时内通过增加细胞增殖来实现。这项研究表明，在肝素存在的情况下，VEGF-A165 是体外诱导 pHEndEC 增殖、血管生成和伤口愈合的有效诱导剂。VEGF-A165 可能是一种重要的有丝分裂原，对于人 BNB 的发育和对周围神经损伤的恢复是必要的。
