AA147对Aβ诱导的阿尔茨海默病细胞模型的作用
Effect of AA147 on a cell model of Alzheimer's disease induced by β-amyloid
王慧 1侯庆明1
作者信息
- 1. 青岛大学神经再生与康复研究院,山东青岛 266071
- 折叠
摘要
目的 探讨AA147能否通过激活内质网应激相关蛋白激活转录因子-6(ATF-6)对阿尔茨海默病细胞模型发挥保护作用.方法 培养孕期为17～19 d的SD大鼠胎鼠皮质神经元,用β-淀粉样蛋白(Aβ)1-42诱导建立阿尔茨海默病细胞模型.实验随机分为对照组(10 μmol/L的二甲基亚砜(DMSO)预处理神经元12 h)、Aβ1-42处理组(2 μmol/L的Aβ1-42诱导神经元24 h)、Aβ1-42+DMSO组(10 μmol/L的DMSO预处理神经元12 h后加入2 μmol/L的Aβ1-42诱导神经元24 h)、Aβ1-42+AA147组(10 μmol/L的AA147预处理神经元12 h后加入2 μmol/L的Aβ1-42诱导神经元24 h).采用CCK-8比色法、免疫荧光染色法检测各组神经元的存活率,Western blot法检测各组神经元中ATF-6及其下游蛋白p-Akt的表达情况.结果 CCK-8检测显示,各组神经元存活率差异有显著意义(F=47.42,P＜0.01),其中Aβ1-42组细胞活力较对照组明显降低(t=9.42,P＜0.05),Aβ1-42+AA147组细胞活力较Aβ1-42组明显升高(t=5.73,P＜0.05).免疫荧光染色结果显示,Aβ1-42组神经元存活数量较对照组显著降低,Aβ1-42+AA147组较Aβ1-42组显著提高,Aβ1-42+DMSO组与Aβ1-42组比较无明显差异.Western blot结果显示,各组ATF-6及p-Akt蛋白表达差异有显著性(F=11.77、26.10,P＜0.05),其中Aβ1-42组ATF-6及p-Akt蛋白表达明显低于对照组,Aβ1-42+AA147组高于Aβ1-42组,差异有统计学意义(P＜0.05),而Aβ1-42+DMSO组与Aβ1-42组比较差异无显著性(P＞0.05).结论 AA147可通过激活ATF-6及其下游信号通路p-Akt的表达发挥对阿尔茨海默病细胞模型的神经保护作用.
Abstract
Objective To investigate whether AA147 can exert a neuroprotective effect on a cell model of Alzheimer's di-sease(AD)by activating the endoplasmic reticulum stress-related protein activating transcription factor 6(ATF6).Methods Cortical neuronsfrom embryonic day 17-19 Sprague-Dawley rat embryos were cultured and induced with β-amyloid(Aβ)1-42 to established a cell model of AD.The cells were divided into control group(neurons pretreated with 10 μmol/L DMSO for 12 h),Aβ1-42 treatment group(neurons induced by 2 μmol/L Aβ1-42 for 24 h),Aβ1-42+DMSO group(neurons pretreated with 10 μmol/L DMSO for 12 h and then induced by 2 μmol/L Aβ1-42 for 24 h),and Aβ1-42+AA147 group(neurons pretreated with 10 μmol/L AA147 for 12 h and then induced by 2 μmol/L Aβ1-42 for 24 h).CCK-8 assay and immunofluorescent staining were used to measure the viabilityof neurons in each group,and Western blot was used to measure the expression of ATF-6 and p-Akt in neurons.Results CCK8 assay showed that there was a significant difference in the viability rate of neurons between groups(F=47.42,P＜0.01);compared with the control group,the Aβ1-42 group had a significant reduction in the viability of neurons(t=9.42,P＜0.05),and compared with the Aβ1-42 group,the Aβ1-42+AA147 group had a significant increase in the viability of neurons(t=5.73,P＜0.05).Immunofluorescent staining showed that compared with the control group,the Aβ1-42 group had a significant reduction in the number of survived neurons,and compared with the Aβ1-42 group,the Aβ1-42+AA147 group had a significant increase in the number of survived neurons,while there was no significant difference between the Aβ1-42+DMSO group and the Aβ1-42 group.Western blot showed that there were significant differences between groups in the protein expression levels of ATF-6 and p-Akt(F=11.77,26.10;P＜0.05);compared with the control group,the Aβ1-42 group had significantly lower pro-tein expression levels of ATF-6 and p-Akt(P＜0.05),and the Aβ1-42+AA147 group had significantly higher expression levels than the Aβ1-42 group(P＜0.05),while there were no significant differences between the Aβ1-42+DMSO group and the Aβ1-42 group(P＞0.05).Conclusion AA147 can exert a neuroprotective effect on the cell model of AD by activating the expression of the ATF-6 and p-Akt signaling pathway.
关键词
阿尔茨海默病/内质网应激/激活转录因子6/神经保护Key words
Alzheimer disease/endoplasmic reticulum stress/activating transcription factor 6/neuroprotection引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据