摘要
目的探讨β-淀粉样肽(Ap)对人SH—SY5Y神经母细胞瘤细胞突触素(syn)、发动蛋白I(DynI)及衔接蛋白180(AP180)表达的影响。方法分别应用0.01、0.1、1、2及5-μmol/LAβ1-42处理SH-SY5Y细胞,对照组细胞不加任何处理。MTT法检测细胞增殖情况,western blotting检测0.5、1μmol/LAμ1-42处理组及对照组细胞Syn、DynI和AP180蛋白表达,RT—PCR检测1μmol/LAβ1-42处理组及对照组细胞Syn、DynI和AP180mRNA表达。结果0.1μmol/LAl3Im处理细胞即可出现细胞增殖率下降,并呈剂量依赖性负性相关关系.与对照组比较差异均有统计学意义(P〈0.05);0.5μmol/LAB1-42处理细胞可见DynI蛋白表达降低,与对照组比较差异有统计学意义(P〈0.05);1μmol/LAβ1-42处理细胞可见Syn、DynI蛋白及mRNA表达均降低,与对照组比较差异均有统计学意义(P〈0.05);但未见AP180蛋白及mRNA表达水平发生改变。结论AB1-42可引起人SH-SY5Y细胞Syn和DynI表达降低,该改变可能与AD发病中学习记忆能力减退有关。
Objective To study the effect of [3-amyloid peptide (Aβ1-42) on expressions of synaptophysin (Syn), dynamin I (Dyn I) and adaptor protein 180 (AP180) in human neuroblastoma SH-SY5Y cells. Methods Human SH-SY5Y neuroblastoma cells were treated with 0.01, 0.1, 1, 2 and 5 μmol/L Aβ1-42, and control cells were given no treatment. MTT [3-(4,5-dimethylthiazol- 2- yl) - 2, 5- diphenyltetrazolium bromide] reduction of the cells was measured by spectrophotometry. The protein levels of Syn, Dyn I and AP180 in SH-SY5Y cells treated with 0.5 and i μmol/L Aβ1-2 were surveyed by Western blotting. The mRNA levels of Syn, Dyn I and AP180 were detected by Real-time PCR in SH-SY5Y cells treated with 1 μmol/L Aβ1-42. Results SH-SY5Y cells showed obviously decreased reduction rates of MTT after exposure to Aβ1-42(0.1 μmol/L) as compared with the controls (P〈0.05), and dose-dependent negative correlation was noted in these SH-SY5Y cells. The protein level of Dyn I in cells treated with 0.5 μmol/L Aβ1-42 was significantly decreased as compared with that in controls (P〈0.05). The protein and mRNA levels of Syn and Dyn I in cells treated with 1 μmol/L Aβ1.42 were obviously decreased as compared with those in controls (P〈0.05), but the levels of AP180 were not changed. Conclusion Aβ1.42 reduces the levels of Syn and Dyn I in SH-SY5Y cells, which might be a mechanism in connection with cognitive deficit of AD.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2011年第8期774-778,共5页
Chinese Journal of Neuromedicine
基金
基金项目:国家自然科学基金(30870986)
贵阳市科学技术计划项目([2009]3-012)
贵阳医学院研究生教育创新基地建设专项经费