秋水仙素诱导的神经元突起溃变对Aβ分泌的影响  

The effect of the degeneration of neuronal neurites induced by colchicine on the secretion of amyloid β-protein

作　　者：陈美婉[1] 罗焕敏[1] 肖飞[1] 邴云云[1] 何利明[1] 

机构地区：[1]暨南大学药学院神经药理研究室,广东广州510632

出　　处：《中国药理学通报》2008年第12期1586-1591,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30672450)

摘　　要：目的研究秋水仙素诱导的体外培养新生小鼠大脑皮层神经元突起溃变对β-淀粉样蛋白(amyloid beta-protein,Aβ)分泌的影响,并初步探讨其可能的机制。方法体外无血清培养新生小鼠大脑皮层神经元,采用NSE免疫细胞化学染色法进行神经元鉴定,选择培养4d的神经元进行实验;通过细胞形态学观察和四唑盐(MTT)比色实验,筛选和确定秋水仙素(colchicine)合适的造模浓度;用ELISA法检测秋水仙素组和溶媒对照组Aβ在不同时间点的分泌量;用RT-PCR测定β淀粉样前体蛋白(APP)、早老素1(PS1)和β位点APP内切酶(BACE)基因的表达。结果细胞形态学观察结果表明,终浓度为0.5mg·L-1的秋水仙素作用4h后,神经元突起明显缩短而胞体形态无变化,同时MTT比色实验测得其吸光度与对照组比差异无显著性,胞体活性没有受到明显影响。ELISA法实验结果表明,神经元突起溃变后,Aβ分泌量在4、8h时升高,与溶媒对照组相比差异有显著性。RT-PCR法表明,APP、PS1和BACE mRNA在4、8h时的表达量较溶媒对照组增加,差异有显著性。结论秋水仙素诱导的神经元突起溃变可增加Aβ分泌,其可能与增加APP,PS1和BACE基因表达有关。Aim To study the effect of the degeneration of neuronal neurites from cerebral cortex of newborn mouse in vitro on the secretion of amyloid β-protein, to investigate the possible mechanism. Method The neurons from cerebral cortex of newborn mouse were cultured in vitro and in a new serum-free medium combination, identified by NSE staining. The 4 days neurons were chosen to be cultured for the experiment. The convert phase microscope and MTT assay were used to decide the proper concentration of colchicine. The ELISA test was used to examine the production of Aβ by the neurons at different time point. The expression of APP, PS1 and BACE mRNA was determined by RT-PCR. Results The results showed that colchicine at 0.5 mg · L^-1 had no obvious effect on the cell survival in the cultured neurons by MTT assay. The ELISA test showed that the production of Aβ4Z by the neurons was significantly increased at 4 h and 8 h after colchicine treatment. The expression of APP, PS1 and BACE mRNA were markedly up-regulated, which was distinctly different from the solvent control group. Conclusion The degeneration of neuronnal neurites induced by colchicine can increase the production of Aβ42. It may be associated with up-regulating the expression of APP, PS1 and BACE mRNA.

关 键 词：神经元突起 Β-淀粉样蛋白 秋水仙素 APP PS1 BACE 

分 类 号：R-332[医药卫生] R322.8