Mechanism of agomelatine alleviating anxiety- and depression-like behaviors in APP/PS1 transgenic mice #br#

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doi:10.12307/2025.323

Abstract

Abstract:
BACKGROUND: Agomelatine is a clinically proven treatment for neuropsychiatric symptoms, such as anxiety and depression. Furthermore, our previous study has demonstrated that agomelatine ameliorates cognitive behaviors, hippocampal synaptic plasticity, and brain pathology in a mouse model of Alzheimer’s disease. However, it remains unclear whether agomelatine can improve anxiety and depression-like behaviors in Alzheimer’s disease model mice.
OBJECTIVE: To investigate the improving effects of agomelatine on anxiety- and depression-like behaviors in APP/PS1 transgenic mice and its underlying molecular mechanisms.
METHODS: (1) Eighteen APP/PS1 transgenic mice were randomly divided into model control group (n=9) and model intervention group (n=9). Another wild-type mice were randomized into control group (n=9) and intervention group (n=9). Model intervention group and intervention group were intraperitoneally injected with 10 mg/kg agomelatine per day for 31 continuous days. Behavioral experiments, including the elevated cross maze and forced swimming tests, and mRNA sequencing of the hippocampus were then performed. (2) Mouse hippocampal neuronal cell lines (HT22) and brain microvascular endothelial cell lines (bEnd.3) were cultured and divided into four groups: blank group without any drug, drug group with 20 µmol/L agomelatine, model group with 10 µmol/L β-amyloid 1-42, and experimental group with 10 µmol/L β-amyloid 1-42+20 µmol /L agomelatine. After 24 hours of incubation, protein expression of S416p-tau and S9p-GSK3β in HT22 cells was detected by immunoblotting, and protein expression of low-density lipoprotein receptor-related protein 1 and glycosylation end-product receptor in bEnd.3 cells was detected by immunoblotting.
RESULTS AND CONCLUSION: In the elevated plus maze test, the time spent in the open arms (P < 0.01) and the entries into open arms (P < 0.05) in the mice of model control group were evidently lower than those in the control group, whereas those were obviously increased in the model intervention group compared with the model control group (P < 0.05). Forced swimming test results showed that the immobile time exhibited a marked increase in the model control group compared with the control group (P < 0.05), but it was significantly decreased in the model intervention group compared with the model control group (P < 0.05). Hippocampal tissue mRNA sequencing showed that agomelatine enhanced the expression of low-density lipoprotein receptor-related protein 1 in the hippocampus of APP/PS1 mice. Western blot analysis revealed that the level of S416p-tau in HT22 cells was higher in the model group than the blank group (P < 0.05), while it was markedly decreased in the experimental group compared with the model group (P < 0.05); the level of S9p-GSK3β in HT22 cells was higher in the drug group than the blank group (P < 0.05) as well as higher in the experimental group than the model group (P < 0.05). Moreover, the expression of low-density lipoprotein receptor-related protein 1 in bEnd.3 cells was higher in the experimental group than the model group (P < 0.05). To conclude, agomelatine can alleviate anxiety- and depression-like behaviors in Alzheimer’s disease mice by promoting the clearance of β-amyloid and phosphorylated tau.

Key words: Alzheimer’s disease, agomelatine, APP/PS1 transgenic mice, anxiety, depression

CLC Number:

Li Tian, Ren Yuhua, Gao Yanping, Su Qiang. Mechanism of agomelatine alleviating anxiety- and depression-like behaviors in APP/PS1 transgenic mice #br#

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[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(6): 1176-1182.

Figures/Tables 4

2.1 动物实验结果 2.1.1 实验动物数量分析 18只APP/PS1转基因小鼠与18只野生型小鼠全部进入结果分析。 2.1.2 阿戈美拉汀明显缓解了APP/PS1转基因小鼠的焦虑样行为在高架十字迷宫测试中，与对照组小鼠相比，模型对照组小鼠开臂探索时间缩短[(26.156±4.315)，(9.333±2.409) s，P < 0.01]，提示9月龄APP/PS1转基因小鼠已伴有焦虑样行为出现；经阿戈美拉汀治疗后，模型干预组小鼠开臂探索时间长于模型对照组[(24.191±3.022)，(9.333±2.409) s，P < 0.01]，见图1A，表明阿戈美拉汀有效改善了APP/PS1转基因小鼠表现出的焦虑样行为。同时，模型对照组小鼠开放臂进入次数少于对照组(2.333±0.441，10.222±1.188，P < 0.001)，模型干预组小鼠开放臂进入次数多于模型对照组(7.667±1.434，2.333±0.441，P < 0.01)，见图1B。这些结果表明，APP/PS1转基因小鼠表现出焦虑样行为，而阿戈美拉汀有效改善了APP/PS1转基因小鼠表现出的焦虑样行为。 2.1.3 阿戈美拉汀有效改善了APP/PS1转基因小鼠的抑郁样行为在强迫游泳测试中，模型对照组小鼠强迫游泳实验后4 min内的不动时间长于对照组[(199.222±9.334)，(108.667±10.603) s，P < 0.001]，提示9月龄APP/PS1转基因小鼠已伴有抑郁样行为出现；模型干预组小鼠强迫游泳实验后4 min内的不动时间短于模型对照组[(137.444±20.626)，(199.222±9.334) s，P < 0.01]，见图2，结果表明阿戈美拉汀可以明显减轻APP/PS1转基因小鼠的抑郁样行为。 2.2 阿戈美拉汀抑制β-淀粉样蛋白1-42诱导的HT22细胞tau蛋白过度磷酸化并促进S9p-GSK3β表达阿尔茨海默病脑内神经元tau过度磷酸化病理与其异常的精神行为表现有关[13-14]。为此，此次实验进一步利用培养的HT22细胞进行了S416p-tau和S9p-GSK3β蛋白表达检测，以检测阿戈美拉汀对β-淀粉样蛋1-42所致神经元tau蛋白磷酸化的影响。如图3所示，模型组HT22细胞的S416p-tau蛋白表达高于空白组(P < 0.05)，实验组HT22细胞的S416p-tau蛋白表达低于模型组(P < 0.05)，表明阿戈美拉汀处理可有效抑制β-淀粉样蛋1-42诱导的HT22细胞tau蛋白过度磷酸化；药物组HT22细胞的S9p-GSK3β蛋白表达高于空白组(P < 0.01)，实验组HT22细胞的S9p-GSK3β蛋白表达高于模型组(P < 0.05)，提示阿戈美拉汀可能是通过促进S9p-GSK3β表达转而抑制β-淀粉样蛋白1-42诱导的海马神经元tau蛋白过度磷酸化，以拮抗β-淀粉样蛋白所致的神经元损伤，从而缓解神经元tau相关的精神行为异常。 2.3 阿戈美拉汀提高bEnd.3细胞LRP1蛋白的表达通过对小鼠海马组织mRNA测序结果进一步分析(图4A)，GO富集分析显示，阿戈美拉汀可以增强APP/PS1转基因小鼠海马区LRP1的蛋白表达。LRP1被认为是分布在血脑屏障上皮细胞上的一种转运受体，参与β-淀粉样蛋白和tau的向外周转运和清除[15-16]。相反，糖基化终产物受体蛋白是表达在血脑屏障上皮细胞上介导外周β-淀粉样蛋白进入脑内的关键受体[17]。因此，为了很好地模拟和检测APP/PS1转基因小鼠脑内微血管内皮细胞LRP1和糖基化终产物受体的蛋白表达情况，利用bEnd.3细胞和免疫印迹手段检测了β-淀粉样蛋白1-42存在的情况下阿戈美拉汀对bEnd.3细胞LRP1和糖基化终产物受体蛋白表达的影响。结果表明，β-淀粉样蛋白1-42处理可诱导bEnd.3细胞LRP1出现下调趋势，同时给予阿戈美拉汀和β-淀粉样蛋白1-42处理的bEnd.3细胞LRP1蛋白表达升高 (P < 0.05)；各组bEnd.3细胞糖基化终产物受体蛋白表达比较差异无显著性意义(P > 0.05)，见图4B-D。结果表明，阿戈美拉汀可能在不影响糖基化终产物受体蛋白表达的情况下诱导阿尔茨海默病小鼠脑内微血管内皮细胞LRP1蛋白表达升高，进而促进阿尔茨海默病脑内β-淀粉样蛋白和tau从中枢向外周转运和清除。"

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