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目的探讨β-神经生长因子(β-NGF)转染的大鼠内皮祖细胞分化功能的变化。方法将人β-NGF重组腺病毒(Ad-EGFP-hβ-NGF组)及其阴性对照(Ad-EGFP组)感染至大鼠内皮祖细胞,同时设置空白对照(NC)组,观察细胞形态学改变,Western blot检测不同组酪氨酸蛋白激酶A(TrkA)表达水平,ELISA法检测不同组培养液中血管内皮生长因子(VEGF)、血管性血友病因子(vWF)及碱性成纤维生长因子(bFGF)水平。结果 Ad-EGFP-hβ-NGF感染至大鼠内皮祖细胞1周后可见部分细胞成球,呈干细胞样改变。Ad-EGFP-hβ-NGF组内皮祖细胞TrKA相对蛋白表达水平(0.67±0.23)明显高于Ad-EGFP组(0.20±0.07)及NC组(0.27±0.12)(P<0.01)。Ad-EGFP-hβ-NGF组培养液中VEGF、vWF及bFGF水平明显高于Ad-EGFP组及NC组(P<0.01)。结论β-NGF通过激活酪氨酸激酶信号通路,增加促血管生长因子的分泌,促进内皮祖细胞的分化,多途径发挥血管修复和再生的功能。
Objective To investigate the changes of differentiation function of rat endothelial progenitor cells transfected with β-nerve growth factor (β-NGF). Methods Human endothelial progenitor cells (EPCs) were infected with Ad-EGFP-hβ-Ad-EGFP and Ad-EGFP and blank control (NC) groups were established. Morphology The expression of tyrosine kinase A (TrkA) in different groups was detected by Western blot and the expressions of vascular endothelial growth factor (VEGF), von Willebrand factor (vWF) and basic fibroblast growth Factor (bFGF) level. Results Ad-EGFP-hβ-NGF infection in rat endothelial progenitor cells after 1 week showed some cells into the ball, showed a change in stem cells. The relative protein expression of TrKA in Ad-EGFP-hβ-NGF group (0.67 ± 0.23) was significantly higher than that in Ad-EGFP group (0.20 ± 0.07) and NC group (0.27 ± 0.12) (P <0.01). The levels of VEGF, vWF and bFGF in Ad-EGFP-hβ-NGF group were significantly higher than that in Ad-EGFP group and NC group (P <0.01). Conclusion β-NGF activates tyrosine kinase signaling, increases the secretion of vascular growth factor, promotes the differentiation of endothelial progenitor cells, and functions in many ways to repair and regenerate blood vessels.