阿托伐他汀对淀粉样β蛋白诱导大鼠原代海马神经元损伤保护作用的研究  

Neuroprotective effect of atorvastatin on amyloid-β protein-induced neurotoxicity in cultured hippocampal neurons

作　　者：王楠[1,2] 隋海娟[1] 张玲玲[1] 金英[1] 赵彬[2] 

机构地区：[1]辽宁医学院药理学教研室,121001 [2]沈阳市第五人民医院,110023

出　　处：《中国神经免疫学和神经病学杂志》2017年第3期178-182,共5页Chinese Journal of Neuroimmunology and Neurology

摘　　要：目的探讨阿托伐他汀(atorvastatin,Ato)对β淀粉样蛋白(Aβ)诱导的体外培养原代海马神经元损伤保护作用。方法选用出生0~24hSprague-Dawley大鼠乳鼠,解剖显微镜下分离海马,进行海马神经元体外培养,培养10d后用于实验。将培养的海马神经元分为3组:(1)正常对照组:加入含有1%(质量浓度)DMSO培养基;(2)Aβ1-42组:加入1.25μmol/L Aβ;(3)Aβ(1.25μmol/L)+不同浓度阿托伐他汀组(0.1、0.5、1、2.5μmol/L)。应用CellTiter-GloTM荧光细胞活性试剂盒检测培养海马神经元ATP含量,用以反映神经元活力;通过CytoTox-ONETM试剂盒测定培养细胞上清液中乳酸脱氢酶(LDH)含量,用以测定神经元细胞膜的损伤程度;应用免疫荧光染色观察神经元突触素(SYP)、突触后致密蛋白95(PSD-95)、bassoon蛋白表达。Western印迹法半定量检测海马神经元突触蛋白SYP、PSD-95、bassoon的改变。结果与正常对照组比较,培养10d海马神经元经1.25μmol/L Aβ1-42作用48h后,其ATP和LDH水平均明显降低(均P<0.01),而0.5、1.0、2.5μmol/L Ato组能够明显抑制Aβ1-42引起的ATP和LDH水平降低(P<0.01)。免疫荧光及Western blot结果显示,1.25μmol/L Aβ1-42可使海马神经元SYP、PSD-95、bassoon蛋白表达明显降低,而Ato能够明显抑制Aβ1-42引起的SYP、PSD-95、bassoon蛋白表达降低。结论 Ato对Aβ诱导的体外培养海马神经元毒性有保护作用,这种保护作用可能与Ato对抗Aβ1-42引起的海马神经元SYP、PSD-95、bassoon表达降低有关。Objective To investigate the neuroprotective effect of atorvastatin（Ato）on amyloid-βprotein-induced neurotoxicity in cultured hippocampal neurons.Methods Primary cultures were obtained from hippocampi of 0hto 24h-old Sprague-Dawley rats.After 10 days in culture,the hippocampi cells were divided into a control group,an Aβ1-42（1.25μmol/L）group,and a treatment group with different Ato concentrations（0.1,0.5,1.0,2.5μmol/L）.The ATP content of the culture was quantified using CellTiter-GloTMassay.The lactate dehydrogenase released into the culture media was measured using CytoTox-ONETMassay.The changes of SYP,bassoon and PSD-95 proteins were observed by immunofluorescence staining.SYP,bassoon and PSD-95 protein were evaluated using Western blotting.Results Compared with the normal control group,in hippocampal neurons cultured for 10 dby Aβ1-42（1.25μmol/L）for 48 h,ATP% and LDH% decreased significantly（P〈0.01）.Adding Ato（0.5,1.0,2.5μmol/L）significantly inhibited Aβ1-42 induced ATP%and LDH% decreases（P〈0.01）.Immunofluorescence and Western results showed that Aβ1-42（1.25μmol/L）decreased SYP,bassoon and PSD-95 protein expression.Compared with the Aβ1-42 group,Ato could significantly inhibit the Aβ1-42 induced above effects.Conclusions Ato has a protective effect against the neurotoxicity of amyloid beta protein to cultured hippocampal neurons in vitro,which may be related to the inhibition of synaptic proteins.

关 键 词：阿尔茨海默病 阿托伐他汀 海马神经元 淀粉样Β蛋白 突触蛋白 

分 类 号：R742.89[医药卫生—神经病学与精神病学]