N-butylphthalide affects cognitive function of APP/PS1 transgenic mice (Alzheimer’s disease model)

doi:10.3969/j.issn.2095-4344.1182

Abstract

Abstract:

BACKGROUND: Amyloid-β protein is a key factor of pathogenesis of Alzheimer’s disease. N-butylphthalide, an anti-cerebral ischemia drug, has been shown to have therapeutic effects in vascular dementia.
OBJECTIVE: To investigate whether N-butylphthalide treating APP/PS1 transgenic mice via the SPEP-ERK-CREB signaling pathway.
METHODS: Thirty male APP/PS1 transgenic mice (mouse model of Alzheimer’s disease), SPF grade, aged 12 months were provided by Beijing Huafukang Biotechnology Co., Ltd., and 10 C57BL/6 wild-type mice, SPF grade, were provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. The animals were divided into four groups (n=10/group): C57BL/6 wild-type mice group, APP/PS1 group, and 10 and 30 mg/kg N-butylphthalide groups. The former two groups were given same volume of vegetable oil via gavage, for 17 consecutive days. The behavioral test was performed at 9-16 days. N-butylphthalide was given 40 minutes prior to Morris water maze task. The mice were killed at 17 days, and the proteins levels of STEP61, pERK1/2 and pCREB in brain cortex were detected by western blot assay.
RESULTS AND CONCLUSION: (1) Consecutive 16-day 10 and 30 mg/kg N-butylphthalide could significantly improve spatial learning disability and working memory disorder of APP/PS1 transgenic mice in Morris water maze test. (2) N-butylphthalide partly reduced the level of activated STEP61, and inhibited the dephosphorylation of ERK and CREB. (3) These results suggest that N-butylphthalide protects against amyloid-β-induced neurodegeneration and cognitive decline in APP/PS1 transgenic mice, which may be potential for Alzheimer’s disease.

Key words: Alzheimer Disease, Amyloid, Cognition

CLC Number:

R496
R318

Zhang Lin1, Liu Jinjie1, Zhao Yan2, Liu Yi1, Lin Jianwen1. N-butylphthalide affects cognitive function of APP/PS1 transgenic mice (Alzheimer’s disease model) [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(在线): 1-6.

Figures/Tables 1

2.1 实验动物数量分析实验选用APP/PS1转基因小鼠30只，C57BL/6野生型小鼠10只，实验过程无脱失，全部进入结果分析。 2.2 丁苯酞可显著改善阿尔茨海默病转基因小鼠空间学习记忆能力见图1。在定位航行实验中，APP转基因小鼠寻找平台的潜伏期明显增加，与同龄野生型小鼠相比较有显著性差异，结果表明APP转基因小鼠对水迷宫学习和记忆的获取能力明显受损，见图1A，B。而两者的平均游泳速度没有明显差异，这表明APP转基因小鼠没有运动能力的缺损，图1E-H。在空间探索实验中，APP转基因小鼠花费在目标象限的时间明显减少，与同龄野生型小鼠相比较有显著性差异，表明APP转基因小鼠对水迷宫空间认知和记忆能力明显受损。在隐蔽平台实验中，APP/PS1 转基因小鼠比野生型小鼠有更长的逃避潜伏期。而两者花费在目标象限的时间在可视平台实验中没有明显的差别，这表明APP转基因小鼠没有视觉能力的缺损。总的来说12月龄APP转基因小鼠表现出明显的海马依赖的学习和记忆能力的缺损，见图1E，F。最后1次训练的24 h后转基因小鼠与野生型相比在目标象限中花费的时间更少。每天给予10和30 mg/kg丁苯酞可以明显改善APP/PS1转基因小鼠学习记忆的缺损。由于有非常显著的药物效应，对逃避潜伏期数据进行了分析。首先，与C57BL/6小鼠相比，APP/PS1小鼠的逃避潜伏期明显受损(P < 0.001)，说明APP/PS1小鼠存在明显学习记忆缺陷；同时，发现10 mg/kg丁苯酞处理的APP/PS1小鼠与C57BL/6小鼠相比，在逃避潜伏期上无显著差异(P < 0.05；见图1F)。说明每天给药10 mg/kg或 30 mg/kg丁苯酞可明显减轻APP/PS1所致的学习障碍。 2.3 丁苯酞显著降低转基因小鼠的STEP61水平 12月龄APP/PS1转基因小鼠的脑皮质突触小体STEP61蛋白水平是同龄野生型小鼠的203%(P < 0.001)。给予APP/PS1小鼠每天10 mg/kg或30 mg/kg丁苯酞灌胃，可检测到STEP61磷酸化比例在皮质突触小体表达量较APP/PS1小鼠增加，尤其是10 mg/kg丁苯酞处理组(P < 0.01，见图2)。2.4 丁苯酞可增加β淀粉样蛋白介导的ERK和CREB磷酸化水平实验检测到APP/PS1转基因小鼠皮质pERK1/2和pCREB水平较正常小鼠明显减少(61.82%，30.5%，P < 0.001)，见图3，4。给予APP/PS1小鼠每天10 mg/kg或 30 mg/kg丁苯酞灌胃后，明显增加了pERKs和pCREB水平，尤其是10 mg/kg丁苯酞处理组(P < 0.05)。作者之前的结果表明STEP61负性调控了β淀粉样蛋白介导的ERK1/2和CREB磷酸化水平和可能的活性(P < 0.01)，提示丁苯酞可以调控STEP/ERK/CREB通路。"

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