加兰他敏抗β淀粉样蛋白的神经保护机制研究  

Protective effects and mechanisms of galantamine on neurotoxicity induced by β-Amyloid protein

作　　者：李倩倩[1] 杨珊珊[1] 黄硕[1] 赵婷婷[1] 孙莉娜[1] 柴丽[1] 

机构地区：[1]哈尔滨医科大学附属第一医院神经内科,黑龙江150001

出　　处：《脑与神经疾病杂志》2017年第2期110-115,共6页Journal of Brain and Nervous Diseases

基　　金：黑龙江省卫计委科研课题(2013007)

摘　　要：目的观察加兰他敏(Gal)对β淀粉样蛋白(Aβ)损伤人神经母细胞瘤细胞(SH-SY5Y)后β淀粉样前体蛋白(APP)代谢通路的影响,探讨Gal的神经保护机制。方法采用5μMAβ_(1-40)作用于SH-SY5Y细胞制备体外细胞损伤模型,0.3μM加兰他敏对Aβ_(1-40)处理的细胞进行干预并与正常细胞进行对照研究。倒置显微镜下观察细胞形态,应用噻唑蓝比色法(MTT)检测细胞活力,Western-blot技术定量检测各组APP,sAPPα,β-淀粉样前体蛋白剪切酶-1(BACE1)表达水平。结果 Aβ_(1-40)孵育细胞24h之后,细胞损伤明显,存活率从95.78.±2.5%降到62.93±2.1%,与对照组相比差异显著(P<0.01);Western-blot显示细胞内BACE1表达增加,APP表达无明显改变,细胞分泌sAPPα降低;在Aβ_(1-40)孵育之前给予加兰他敏作用24h,细胞损伤程度减轻,细胞的存活率上升(85.26±5.3%)(P<0.01),细胞内BACE1表达较Aβ组下降,APP表达无明显改变,细胞分泌sAPPα升高。结论加兰他敏通过抑制Aβ_(1-40)诱导的APP的异常代谢发挥神经保护作用。Objective To observe the effects of galantamine （ Gal ） on APP metabolic pathways in SH- SY5Y cells treated with amyloid- β peptide （ Aβ） , and explore the mechanism underlying the protective effect of Gal against Aβ -induced neurotoxity. Method We set up a ceil moldel through using SH-SY5Y ceils incubated with A β 1-40 （ 5 μL mol · L^-1 ） for 24h, which is also pretreated with Gal （ 0.3μL mol · L^-1 ） .when investgating the protective effects and mechanisms of Gal against A β -induced neurotoxity. Morphological changes were observed using macroscope and cell viability was determined by MTT assay. The amount of sAPP α released in the cell culture media ,and also the expression of cellular APP and BACE1 was determined by Western-blotting. Results Gal pretreatment significantly prevented A β1-40-induced cell death. Cell viability was significantly decreased in cultures treated with 5 μL mol · L^-1A β 1-40 for 24 hours （ 62.93 ± 2.1% ） compared with the control group （ 95.78± 2.5% ） （ P〈0.01 ） ;A β 1-40-indueed toxicity was significantly prevented by pre-ineubation of cells with 0.3 μL mol · L^-1 Gal for 24h, since cell viability increased to 85.26 ＋ 5.3% （ P〈0.01 ） .Both the overexpression of BACE1 and decrease of sAPPct were observed in A β 1-40-damaged cells. However, all these alterations were found to be reversed by Gal pretreatment. No effect on cellular APP levels was found in both A β 1-40 group and Ga] group （ P〉0.05 ） .Conclusions Gal protect against the neuronal damage induced by A β1-40 through reduced cleavage of APP via the β -secretase pathway ,which related to the inhibition of BACE1 expression.

关 键 词：加兰他敏 Β淀粉样蛋白 人神经母细胞瘤细胞 Β淀粉样前体蛋白 淀粉样前体蛋白 β位分解酶 

分 类 号：R741[医药卫生—神经病学与精神病学]