硫必利对Aβ25-35诱导Neuro-2a细胞损伤的保护作用

Neuroprotective effect of thiopride on Aβ25-35 induced toxicity in Neuro-2a cells

目的探讨硫必利对β-淀粉样蛋白Aβ25-35诱导小鼠脑神经母细胞瘤(Neuro-2a)损伤的保护作用及其潜在机制。方法将细胞实验分为7组:空白对照组、模型组和第1实验组至第5实验组。用Aβ25-35(100μmol·L^-1)处理细胞,作为模型组。第1实验组至第5实验组是在模型组基础上,加入5个不同质量浓度(3,10,30,100,300 ng·mL^-1)硫必利处理24 h。用MTT法测定细胞活力,以蛋白质印迹法测定磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/叉头框蛋白O转录因子3(FoxO3)信号传导蛋白的磷酸化情况(灰度值)。结果空白对照组、模型组和第1实验组至第5实验组的细胞活力分别为(100.00±4.00)%,(54.57±4.24)%,(59.26±8.51)%,(64.17±7.84)%,(73.20±7.04)%,(94.52±12.03)%和(98.31±11.24)%。第3实验组至第5实验组与模型组比较,细胞活力均明显升高,差异均有统计学意义(均P<0.05)。空白对照组、模型组和第4实验组的p-FoxO3蛋白表达水平分别为1.05±0.13,1.42±0.18和8.57±3.42;这3组的p-AktThr473蛋白表达水平分别为1.00±0.11,1.31±0.23和1.58±0.26;第4实验组与模型组相比,p-FoxO3、p-AktThr473蛋白表达差异均有统计学意义(均P<0.05)。结论硫必利通过激活PI3K/Akt/Foxo3通路来对抗Aβ25-35在Neuro-2a中的凋亡作用。

Objective To investigate the neuroprotective effect of thiopride on Aβ25-35 induced toxicity in Neuro-2a cells and its potential mechanism.Methods The Neuro-2a cells were divided into 7 groups:blank control group,model group and experimental-1,-2,-3,-4,-5 groups.Except the blank control group,the cells in other groups were treated with Aβ25-35(100μmol·L^-1)as the model group.Then,experimental-1,-2,-3,-4,-5 groups were given 3,10,30,100,300 ng·mL^-1 thiopride,respectively,for 24 h.The cell viability was measured by MTT assay.While expression(gray value)of the phosphorylation of signaling proteins,such as phosphoinositide 3-kinase inhibitor(PI3K)/protein kinase B(Akt)/Forkhead box O3(FoxO3)were determined by Western blotting.Results The cell viability in blank control group,model group and experimental-1,-2,-3,-4,-5 groups were(100.00±4.00)%,(51.57±4.24)%,(59.26±8.51)%,(64.17±7.84)%,(73.20±7.04)%,(94.52±12.03)%,and(98.31±11.24)%,respectively.Compared between experimental-3,-4,-5 groups and model group,the difference were significant(all P<0.05).The expression levels of p-FoxO3 protein in blank control group,model group and the experimental-4 group were 1.05±0.13,1.42±0.18,and 8.57±3.42,respectively;the expression levels of p-AktThr473 were 1.00±0.11,1.31±0.23,and 1.58±0.26,respectively.Compared between experimental-4 group and model group,the differences of the factors were significant(all P<0.05).Conclusion Thiopride can activate the PI3K/Akt/FoxO3 pathway to oppose the apoptotic action of Aβ25-35 in Neuro-2a cells.