基础医学与临床 ›› 2011, Vol. 31 ›› Issue (6): 688-693.

• 研究论文 • 上一篇    下一篇

Aβ1-42对SH-SY5Y细胞T514位点磷酸化CRMP2表达的影响

朱天瑞1,李晓红2,王敏2,张媛媛2,王栋2   

  1. 1. 山东大学医学院
    2. 山东大学附属济南市中心医院
  • 收稿日期:2010-12-02 修回日期:2011-03-14 出版日期:2011-06-05 发布日期:2011-06-06
  • 通讯作者: 李晓红 E-mail:lxh5231@Yahoo.com.cn
  • 基金资助:
    他汀类药物通过Rho/ROCK途径对老年性痴呆β淀粉样蛋白沉积的影响

Effect of Aβ1-42 on expression of T514 phosphorated CRMP2 in SH-SY5Y cells

Tian-rui ZHU1,Xiao-hong LI2,Min WANG3,Yuan-yuan ZHANG3,Dong WANG3   

  1. 1. Medical College, Shandong University
    2. Jinan Central Hospital Affilited to Shandong University
    3.
  • Received:2010-12-02 Revised:2011-03-14 Online:2011-06-05 Published:2011-06-06
  • Contact: Xiao-hong LI E-mail:lxh5231@Yahoo.com.cn

摘要: 目的:探讨β淀粉样蛋白1-42(Aβ1-42)对神经成廇细胞SH-SY5Y轴突生长状态、细胞微管结构及T514位点磷酸化脑衰蛋白反应调节蛋白2 (CRMP2)表达的影响。方法:用全反式维甲酸诱导SH-SY5Y细胞,细胞外给予聚集态Aβ1-42致细胞损伤;用MTT法检测细胞存活率;用细胞化学法测量细胞轴突长度;免疫荧光法观察细胞微管结构及T514位点磷酸化CRMP2的分布;Westen blot分析T514位点磷酸化CRMP2在细胞内的表达。结果:0.1与1μmol/L聚集态Aβ1-42使诱导后SH-SY5Y细胞存活率显著减低(p<0.01);以1μmol/LAβ1-42处理后,细胞轴突长度缩短(p<0.05)。0.1和1μmol/L Aβ1-42处理后细胞微管结构模糊,T514位点磷酸化CRMP2在细胞内荧光分布面积扩大(p<0.05),强调度增加(p<0.01);用0.1和1μmol/L聚集态Aβ1-42处理后细胞内T514位点磷酸化CRMP2表达量显著增加(p<0.01)。 结论:Aβ1-42可增加SH-SY5Y细胞内T514位点磷酸化CRMP2的表达,损害微管结构,抑制轴突生长。

关键词: β淀粉样蛋白 , 神经成瘤细胞, 全反式维甲酸

Abstract: Objective To investigate the effect of β-amyloid peptide 1-42( Aβ1-42) on neurite outgrowth, microtubule structure and expression of T514 phosphorated collapsin response mediator protein 2 (CRMP2) in SH-SY5Y cells. Methords Making cells model by adding aggregated Aβ1-42 in the culture medium of all-trans retinoic acid induced neuroblastoma cells SH-SY5Y. MTT methord was employed to identify the viability of SH-SY5Y cells. Cytochemistry was used to measure neurite length of induced SH-SY5Y cells. The intracellular microtubule structure and distribution of T514 phosphorated CRMP-2 was detected by immunofluorescence . Westen blot was used to detect the influence of Aβ1-42 on total expression of T514 phosphorated CRMP-2 . Results The viability of SH-SY5Y showed significant decrease in response to 24h aggregated Aβ1-42 exposure at 0.1 and 1μmol/L (p<0.01). Neurites measurement showed that 1μmol/L aggregated Aβ1-42 lead to obvious neutrite retraction (p<0.05). Immunofluorescence showed the CRMP2 immuno-positive expression was higher in both experimental groups (0.1 and 1μmol/L ) (p<0.01) . Westen blot showed the totle expression of T514 phosphorated CRMP-2 in both experimental groups (0.1 and 1μmol/L ) maintain significant higher level compared with control (p<0.01). Conclusion Aβ1-42 can increase intracellular expression of T514 phosphorated CRMP-2 and have a negtive effect on microtubule structure and neurite outgrowth.

Key words: β-amyloid protein , SH-SY5Y, all-trans retinonic acid