来自:中国糖尿病杂志 编辑:王中京 赵湜 丁胜 毛红 牛力 孙敏娴|点击数:|2013-09-26
·糖尿病基础研究·
【摘要】 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)及其阴性对照(Ad-EGFP)感染至大鼠内皮祖细胞,同时设置空白对照(Blank),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感染组内皮祖细胞TrKA相对蛋白表达水平(0.67±0.23)明显高于阴性对照组(0.20±0.07)及空白对照组(0.27±0.12)(P <0.01)。Ad-EGFP-hβ-NGF组培养液中VEGF、vWF及bFGF水平明显高于阴性对照组及空白对照组 (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。
【关键词】 β-神经生长因子;内皮祖细胞;分化;大鼠
THE INFLUENCE OF HUMAN β-NERVE GROWTH FACTOR ON THE DIFFERENCIATION OF ENDOTHELIAL PROGENITOR CELLS WANG Zhong-jing,ZHAO Shi,DING Sheng,et al. Department of Endocrinology, Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Hubei 430014, China
【Abstract】 Objective To evaluate the influence of β-nerve growth factor(β-NGF) on the differentiation of Endothelial Progenitor Cells (EPCs). Methods Primary rat EPCs were isolated from rat bone marrow. The purified EPCs were divided into three groups, and they were infected with human β-NGF adenovirus(Ad-EGFP-hβ-NGF) and its control(Ad-EGFP) respectively, an addition pseudo-infection was acted as blank control. 48 hours after infection, we observed and compared their morphological changes. The TrkA expression was determined by western blot, and we further detected the secretion of VEGF, vWF and bFGF in EPCs by ELISA. Results We found that EPCs were originally identified as fibroblastic appearance and then appeared to be epithelial-like. Ad-EGFP-hβ-NGF infection induced EPCs a sphere-like growth, up-regulated its expression of TrkA (P <0.01), and enhanced the secretion of VEGF, vWF and bFGF (P <0.01) significantly compared to the blank and Ad-EGFP groups. Conclusions β-NGF activates TrkA signaling pathway, and up-regulates its secretion of VEGF, vWF and bFGF, which facilitate the differentiation of EPCs and contribute to angiopoiesis or vascular repair.
【Key words】β-Nerve growth factor; Endothelial progenitor cell; Differentiation; rat
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