葛根素减轻APP/PS1双转基因小鼠的认知障碍和tau蛋白过磷酸化
|
摘要
该实验利用APP/PS1双转基因模型小鼠,观察葛根素对小鼠学习记忆能力及tau蛋白磷酸化的影响。选用APP/PS1双转基因模型小鼠,3个月龄后开始灌胃给药。给药3个月后,通过Morris水迷宫实验测试小鼠学习记忆能力的变化;行为学测试结束后,处死动物,留取脑组织标本,ELISA法检测皮层Aβ水平;Western blot法检测皮层tau蛋白、磷酸化tau蛋白、GSK3β及磷酸化GSK3β(Ser9)蛋白表达情况。Morris水迷宫实验显示,APP/PS1双转基因模型小鼠逃避潜伏期较正常对照组明显延长,原象限停留时间明显缩短。葛根素组小鼠逃避潜伏期较模型组明显缩短,原象限停留时间明显延长。与正常对照组比较APP/PS1双转基因模型小鼠皮层Aβ水平增多,磷酸化tau蛋白表达显著增加,磷酸化GSK3β(Ser9)蛋白表达降低;给予葛根素后,APP/PS1转基因小鼠寻台潜伏期明显缩短,Aβ水平减少,磷酸化tau蛋白表达显著减少,磷酸化GSK3β(Ser9)蛋白表达增加。葛根素通过减少Aβ的生成,激活GSK3β信号通路,抑制tau蛋白磷酸化,改善APP/PS1双转基因模型小鼠的学习记忆损害。
To observe the effect of puerarin on learning and memory function and tau phosphorylation in APP/PS1 transgenic mice,drugs were administered to 3-month old APP / PS1 transgenic mice. Learning and memory function of mice were assessed by Morris water maze test 3 months after treatment. Animals were decapitated after behavioral test. The levels of Aβ were detected by ELISA,the expression of protein [tau,phosphorylated tau,GSK3β and p-GSK3β( Ser9) ] were assessed by Western blot. Morris water maze test showed that the escape latency of APP / PS1 double transgenic mice was significantly longer than that of the normal control group,and the residence time of the original quadrant was significantly shorter. The escape latency of puerarin group was significantly shorter and the residence time of the original quadrant was prolonged compared with the model group. Compared with the normal control group,the levels of Aβ in the cortex of APP / PS1 transgenic mice were increased,the expression of phosphorylated tau was significantly increased,and the expression of phosphorylated GSK3β( Ser9) protein was decreased. Treatment with puerarin,the latency of APP / PS1 transgenic mice was significantly reduced,the level of Aβ was decreased,the expression of phosphorylated tau was significantly decreased,and the expression of phosphorylated GSK3β( Ser9) protein was increased. Puerarin improves the learning and memory impairment by reducing the formation of Aβ,activating the GSK3β signaling pathway,inhibiting the phosphorylation of tau in APP / PS1 double transgenic mice.
To observe the effect of puerarin on learning and memory function and tau phosphorylation in APP/PS1 transgenic mice,drugs were administered to 3-month old APP / PS1 transgenic mice. Learning and memory function of mice were assessed by Morris water maze test 3 months after treatment. Animals were decapitated after behavioral test. The levels of Aβ were detected by ELISA,the expression of protein [tau,phosphorylated tau,GSK3β and p-GSK3β( Ser9) ] were assessed by Western blot. Morris water maze test showed that the escape latency of APP / PS1 double transgenic mice was significantly longer than that of the normal control group,and the residence time of the original quadrant was significantly shorter. The escape latency of puerarin group was significantly shorter and the residence time of the original quadrant was prolonged compared with the model group. Compared with the normal control group,the levels of Aβ in the cortex of APP / PS1 transgenic mice were increased,the expression of phosphorylated tau was significantly increased,and the expression of phosphorylated GSK3β( Ser9) protein was decreased. Treatment with puerarin,the latency of APP / PS1 transgenic mice was significantly reduced,the level of Aβ was decreased,the expression of phosphorylated tau was significantly decreased,and the expression of phosphorylated GSK3β( Ser9) protein was increased. Puerarin improves the learning and memory impairment by reducing the formation of Aβ,activating the GSK3β signaling pathway,inhibiting the phosphorylation of tau in APP / PS1 double transgenic mice.
引文
[1]Miller E C,Teravskis P J,Dummer B W,et al.Tau phosphorylation and tau mislocalization mediate soluble Aβoligomer-induced AMPA glutamate receptor signaling deficits[J].Eur J Neurosci,2014,39(7):1214.
[2]Ittner L M,Ke Y D,Delerue F,et al.Dendritic function of tau mediates amyloid-beta toxicity in Alzheimer's disease mouse models[J].Cell,2010,142(3):387.
[3]Zeng Q,Zheng M,Zhang T,et al.Hippocampal neurogenesis in the APP/PS1/nestin-GFP triple transgenic mouse model of Alzheimer's disease[J].Neuroscience,2016,314:64.
[4]Zhou Y,Xie N,Li L,et al.Puerarin alleviates cognitive impairment and oxidative stress in APP/PS1 transgenic mice[J].Int J Neuropsychopharmacol,2014,17(4):635.
[5]吴佳莹,羊红玉,赵青威,等.抗阿尔茨海默病的新靶点及药物研发进展[J].中国医院药学杂志,2013,33(19):1615.
[6]Zhao S S,Yang W N,Jin H,et al.Puerarin attenuates learning and memory impairments and inhibits oxidative stress in STZ-induced SAD mice[J].Neurotoxicology,2015,51:166.
[7]马涛,辛文峰,张文生,等.三七皂苷R1对Aβ1-42诱导的SHSY5Y细胞凋亡的保护作用[J].中国中药杂志,2015,40(2):303.
[8]Zhou D,Liu H,Li C,et al.Atorvastatin ameliorates cognitive impairment,Aβ1-42production and Tau hyperphosphorylation in APP/PS1 transgenic mice[J].Metab Brain Dis,2016,31(3):369.
[9]梅峥嵘,谭湘萍,黄汉辉,等.葛根素对阿尔茨海默病细胞模型Aβ蛋白的抑制作用[J].中国现代应用药学杂志,2015,32(1):5.
[10]Quintanilla R A,von Bernhardi R,Godoy J A,et al.Phosphorylated tau potentiates Aβ-induced mitochondrial damage in mature neurons[J].Neurobiol Dis,2014,71:260.
[11]Yang C C,Kuai X X,Gao W B,et al.Morroniside-induced PP2A activation antagonizes Tau hyperphosphorylation in a cellular model of neurodegeneration[J].J Alzheimers Dis,2016,51(1):33.
[12]Oliveira J M,Henriques A G,Martins F,et al.Amyloid-βmodulates both AβPP and Tau phosphorylation[J].J Alzheimers Dis,2015,45(2):495.
[13]Hong X P,Chen T,Yin N N,et al.Puerarin ameliorates D-galactose induced enhanced hippocampal neurogenesis and tau hyperphosphorylation in rat brain[J].J Alzheimers Dis,2016,51(2):605.
[14]Li L,Liu Z,Liu J,et al.Ginsenoside Rd attenuates beta-amyloid-induced tau phosphorylation by altering the functional balance of glycogen synthase kinase 3beta and protein phosphatase2A[J].Neurobiol Dis,2013,54:320.
[2]Ittner L M,Ke Y D,Delerue F,et al.Dendritic function of tau mediates amyloid-beta toxicity in Alzheimer's disease mouse models[J].Cell,2010,142(3):387.
[3]Zeng Q,Zheng M,Zhang T,et al.Hippocampal neurogenesis in the APP/PS1/nestin-GFP triple transgenic mouse model of Alzheimer's disease[J].Neuroscience,2016,314:64.
[4]Zhou Y,Xie N,Li L,et al.Puerarin alleviates cognitive impairment and oxidative stress in APP/PS1 transgenic mice[J].Int J Neuropsychopharmacol,2014,17(4):635.
[5]吴佳莹,羊红玉,赵青威,等.抗阿尔茨海默病的新靶点及药物研发进展[J].中国医院药学杂志,2013,33(19):1615.
[6]Zhao S S,Yang W N,Jin H,et al.Puerarin attenuates learning and memory impairments and inhibits oxidative stress in STZ-induced SAD mice[J].Neurotoxicology,2015,51:166.
[7]马涛,辛文峰,张文生,等.三七皂苷R1对Aβ1-42诱导的SHSY5Y细胞凋亡的保护作用[J].中国中药杂志,2015,40(2):303.
[8]Zhou D,Liu H,Li C,et al.Atorvastatin ameliorates cognitive impairment,Aβ1-42production and Tau hyperphosphorylation in APP/PS1 transgenic mice[J].Metab Brain Dis,2016,31(3):369.
[9]梅峥嵘,谭湘萍,黄汉辉,等.葛根素对阿尔茨海默病细胞模型Aβ蛋白的抑制作用[J].中国现代应用药学杂志,2015,32(1):5.
[10]Quintanilla R A,von Bernhardi R,Godoy J A,et al.Phosphorylated tau potentiates Aβ-induced mitochondrial damage in mature neurons[J].Neurobiol Dis,2014,71:260.
[11]Yang C C,Kuai X X,Gao W B,et al.Morroniside-induced PP2A activation antagonizes Tau hyperphosphorylation in a cellular model of neurodegeneration[J].J Alzheimers Dis,2016,51(1):33.
[12]Oliveira J M,Henriques A G,Martins F,et al.Amyloid-βmodulates both AβPP and Tau phosphorylation[J].J Alzheimers Dis,2015,45(2):495.
[13]Hong X P,Chen T,Yin N N,et al.Puerarin ameliorates D-galactose induced enhanced hippocampal neurogenesis and tau hyperphosphorylation in rat brain[J].J Alzheimers Dis,2016,51(2):605.
[14]Li L,Liu Z,Liu J,et al.Ginsenoside Rd attenuates beta-amyloid-induced tau phosphorylation by altering the functional balance of glycogen synthase kinase 3beta and protein phosphatase2A[J].Neurobiol Dis,2013,54:320.