上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

ERK/CREB信号通路在异菝葜皂苷元促进Aβ损伤SH-SY5Y细胞BDNF表达中的作用

张瑞1, 2，夏志明1，孙小余1，李加梅1，张永芳1, 2，胡雅儿1, 2   

  1. 上海交通大学 医学院 1．细胞调控研究室，2．转化医学协同创新中心，上海 200025
  • 出版日期:2016-05-28 发布日期:2016-05-26
  • 通讯作者: 胡雅儿, 电子信箱: yaerhu@shsmu.edu.cn; 张永芳, 电子信箱: zhangyongfang1@yahoo.com。
  • 作者简介:张瑞（1981—）， 女， 实验师， 博士; 电子信箱: rzhang0621@163.com。
  • 基金资助:

    上海市卫生局科研计划课题资助项目（20124y007）；国家自然科学基金（81573401）

Role of ERK/CREB signal pathway in promotion of BDNF expression in Aβ intoxicated SH-SY5Y cells by smilagenin

ZHANG Rui1, 2, XIA Zhi-ming1, SUN Xiao-yu1, Li Jia-mei1, ZHANG Yong-fang1, 2, HU Ya-er1, 2   

  1. 1.Research Laboratory of Cell Regulation, 2.Collaborative Innovation Center for Translational Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2016-05-28 Published:2016-05-26
  • Supported by:

    Shanghai Municipal Health Bureau Foundation，20124y007；National Natural Science Foundation of China， 81573401

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摘要：

目的 探讨细胞外信号调节激酶/cAMP反应元件结合蛋白（ERK/CREB）信号通路在异菝葜皂苷元（SMI）促进β-淀粉样肽（Aβ）损伤SH-SY5Y细胞脑源性神经营养因子（BDNF）表达中的作用及其机制。方法 建立Aβ损伤SH-SY5Y细胞模型，采用RT-PCR法检测BDNF mRNA的表达，用Western blotting的方法检测pCREB和pERK的变化，最后通过阻断实验确认ERK/CREB信号通路在SMI促进BDNF mRNA表达中的作用。结果 SMI能促进Aβ损伤SH-SY5Y细胞BDNF mRNA的表达（P=0.000），并能上调信号分子CREB和ERK1/2的磷酸化水平（P=0.001，P=0.000）。用RNA干扰的方法阻断CREB的表达后，SMI促进BDNF mRNA表达的作用完全消失;用U0126阻断ERK1/2磷酸化后，SMI促进pCREB水平升高的作用完全消失。结论 SMI通过ERK/CREB通路促进Aβ损伤SH-SY5Y细胞BDNF mRNA的表达。

关键词: 异菝葜皂苷元, 阿尔茨海默症, 脑源性神经营养因子, cAMP 反应元件结合蛋白, 细胞外信号调节激酶1/2

Abstract:

Objective To investigate the role of extracellular signal regulated kinase/cAMP response element binding protein (ERK/CREB) signaling pathway in promotion of brain derived neurotrophic factor (BDNF) expression in β-amyloid (Aβ) intoxicated SH-SY5Y cells by smilagenin (SMI) and relevant mechanisms. Methods The Aβ intoxicated cell model was built and RT-PCR assay was employed to detect the BDNF mRNA expression. Western blotting was used to detect changes in pCREB and pERK expressions. Finally, the role of ERK/CREB signaling pathway in the promotion of BDNF mRNA expression by SMI was confirmed by the use of blocking assay. Results SMI promoted the BDNF mRNA expression in Aβ intoxicated SH-SY5Y cells (P=0.000) and up-regulated the phosphorylation level of signal molecules CREB and ERK1/2 (P=0.001, P=0.000). After blocking the CREB expression by RNA interference, the effect of SMI on promoting the BDNF mRNA expression completely vanished. After blocking the ERK1/2 phosphorylation with U0126, the effect of SMI on promoting the increase in pCREB level completely vanished. Conclusion SMI promotes the BDNF mRNA expression in Aβ intoxicated SH-SY5Y cells through the ERK/CREB pathway.

Key words: smilagenin, Alzheimers disease, brain-derived neurotrophic factor, cAMP response element binding protein, extracellular signal-regulated kinase