中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (17): 2684-2689.doi: 10.3969/j.issn.2095-4344.1709

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

不同剂量血管内皮生长因子在低氧环境下可保护神经干细胞

周经霞，陈 琳，陈擘璨，邢槐杰，闫丽敏，曾超胜，吴海荣，黄裕盛，陈接桂   

  1. 海南医学院第二附属医院神经内科，海南省海口市 570031
  • 修回日期:2019-01-16 出版日期:2019-06-18 发布日期:2019-06-18
  • 通讯作者: 陈琳，硕士，主治医师，海南医学院第二附属医院神经内科，海南省海口市 570031
  • 作者简介:周经霞，女，1979年生，海南省海口市人，2004年海南医学院毕业，副主任医师，主要从事神经内科的临床科研工作。
  • 基金资助:

    海南省卫生计生行业科研项目(1801032054A2011)，项目负责人：周经霞

Protective effects of different doses of vascular endothelial growth factors on neural stem cells under hypoxic environment

Zhou Jingxia, Chen Lin, Chen Bocan, Xing Huaijie, Yan Limin, Zeng Chaosheng, Wu Hairong, Huang Yusheng, Chen Jiegui   

  1. Department of Neurology, the Second Affiliated Hospital of Hainan Medical University, Haikou 570031, Hainan Province, China
  • Revised:2019-01-16 Online:2019-06-18 Published:2019-06-18
  • Contact: Chen Lin, Master, Attending physician, Department of Neurology, the Second Affiliated Hospital of Hainan Medical University, Haikou 570031, Hainan Province, China
  • About author:Zhou Jingxia, Associate chief physician, Department of Neurology, the Second Affiliated Hospital of Hainan Medical University, Haikou 570031, Hainan Province, China
  • Supported by:

    the Scientific Research Project of Hainan Provincial Commission of Health and Family Planning, China, No. 1801032054A2011 (to ZJX)

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摘要：

文章快速阅读：

文题释义：
血管内皮生长因子：具有促进血管通透性增加、细胞外基质变性、血管内皮细胞迁移、增殖和血管形成等作用，有5种不同的亚型。
低氧细胞模型：是指由于细胞得不到充足的氧或不能充分利用氧，导致组织、细胞代谢、功能和形态结构发生异常变化的病理过程。

 

摘要
背景：血管内皮生长因子是一种可以促进血管形成的细胞因子，近来有动物实验及临床研究显示其在急性缺血性卒中以及脊髓损伤组织中表达增加，可抑制细胞凋亡，但具体作用机制尚不明确。
目的：以不同剂量血管内皮生长因子干预低氧环境诱导的神经干细胞，观察其保护作用，并探索其作用机制。
方法：分离培养胎鼠大脑皮质神经干细胞，随机分为空白对照组、缺氧组以及低、中、高剂量血管内皮生长因子组。空白对照组在体积分数35%氧气环境培养24 h，缺氧组以及低、中、高剂量血管内皮生长因子组在体积分数3%氧气环境培养24 h，低、中、高剂量血管内皮生长因子组培养基中分别加入20，40和80 μmol/L血管内皮生长因子。CCK-8法、TUNEL法检测各组细胞增殖和凋亡情况，Western blot检测神经细胞凋亡相关蛋白的表达水平。
结果与结论：与空白对照组相比，缺氧组神经干细胞增殖能力下降，凋亡细胞数量明显增加，且细胞中Bcl-2和核因子κB表达水平下降，Bax和TWIK相关的酸敏感钾离子通道蛋白表达水平增加，而不同剂量的血管内皮生长因子都能抑制上述现象，其中40 μmol/L血管内皮生长因子的作用效果最为显著。结果说明血管内皮生长因子能通过干预凋亡相关蛋白表达而影响细胞凋亡，进而发挥神经保护作用，且40 μmol/L血管内皮生长因子效果最强。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0001-8799-966X(周经霞)

关键词: 神经干细胞, 低氧环境, 3%氧气环境, 血管内皮生长因子, 凋亡相关蛋白, 细胞凋亡, 神经保护

Abstract:

BACKGROUND: Vascular endothelial growth factor (VEGF) is a cytokine which can promote angiogenesis. Recently, some animal experiments and clinical studies have shown that VEGF expression is increased in acute ischemic stroke and spinal cord injury, which can inhibit cell apoptosis, but the mechanism is not clear.
OBJECTIVE: To intervene in hypoxic-induced neural stem cells with different doses of VEGF, and to explore the protective effects and mechanism of VEGF.
METHODS: Neural stem cells was obtained from the cerebral cortex of a fetal rat, which were randomly divided into blank control group, hypoxia group and low-, medium- and high-dose VEGF groups. The cells in the blank control group were cultured for 24 hours under 35% oxygen. The cells in the other four groups were cultured for 24 hours under 3% oxygen. In the low-, medium- and high-dose VEGF groups, 20, 40 and 80 μmol/L VEGF were supplemented into the medium, respectively. The proliferation and apoptosis of the cells were detected by cell counting kit-8 method and TUNEL method. The expression level of nerve cell apoptosis related proteins was detected by western blot.
RESULTS AND CONCLUSION: Compared with the blank control group, the proliferation of neural stem cells decreased, the number of apoptotic neural stem cells was increased significantly, the expression levels of Bcl-2 and nuclear factor κB in the cells were decreased, and the expression levels of Bax and TWIK were increased in the hypoxia group. VEGF at all the three doses could inhibit these phenomena, and 40 μmol/L VEGF showed the best efficacy. These results indicate that VEGF can affect cell apoptosis by interfering with the expression of apoptosis-related proteins, and then exert neuroprotective effects, especially at 40 μmol/L.

Key words: neural stem cells, hypoxic environment, vascular endothelial growth factor, apoptosis-related proteins, apoptosis, neuroprotection

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