摘要
目的探讨β淀粉样蛋白1-42(Aβ1-42)对成神经瘤细胞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/L Aβ1-42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。
Objective To investigate the effect of β-amyloid peptide 1-42（Aβ1-42） on neurite outgrowth,microtubule structure and expression of T514 phosphorylated collapsin response mediator protein 2（CRMP2） in SH-SY5Y cells.Methods To develope 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 phosphorylated CRMP-2 was detected by immunofluorescence.Westen blot was used to detect the influence of Aβ1-42 on total expression of T514 phosphorylated CRMP-2.Results The viability of SH-SY5Y showed significant decrease in response to 24h aggregated Aβ1-42 exposure at 0.1 and 1 μmol/L（P0.01）.Neurites measurement showed that 1 μmol/L aggregated Aβ1-42 lead to obvious neutrite retraction（P0.05）.Immunofluorescence showed the CRMP2 immuno-positive expression was higher in both experimental groups（0.1 and1 μmol/L）（P0.01）.Westen blot showed the totle expression of T514 phosphorylated CRMP-2 in both experimental groups（0.1 and 1 μmol/L） maintained significant higher level compared with control（P0.01）.Conclusion Aβ1-42 can increase intracellular expression of T514 phosphorylated CRMP-2 and have a negtive effect on microtubule structure and neurite outgrowth.
出处
《基础医学与临床》
CSCD
北大核心
2011年第6期688-693,共6页
Basic and Clinical Medicine
基金
山东省自然科学基金(Y2008C103)
