【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。更多还原

【Abstract】 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.更多还原