摘要
目的 探究5×FAD小鼠中神经递质动态变化与β-淀粉样蛋白(amyloid-beta, Aβ)沉积及神经炎症的时序关联。方法 选取1.5月龄(病理早期)、3月龄(病理中期)和6月龄(病理晚期)的5×FAD小鼠及C57BL/6J(WT)小鼠,建立阿尔茨海默病(Alzheimer’s disease, AD)进展的时间窗模型。每个年龄点及基因型组别均使用8只小鼠。通过Y迷宫、新物体识别及旷场进行行为学表型量化。利用免疫荧光检测海马和前额叶皮层(prefrontal cortex, PFC)的Aβ表达和胶质细胞激活水平,苏木素-伊红(HE)染色观察海马神经元形态,并采用高效液相色谱法测定海马和PFC的神经递质水平。结果 研究发现,1.5月龄AD模型组小鼠海马中γ-氨基丁酸水平较WT组小鼠下降;与WT组相比,3月龄AD模型组小鼠海马部分神经递质如乙酰胆碱、谷氨酸、5-羟色胺和5-羟吲哚乙酸水平发生显著变化,此时海马和PFC已有少量Aβ沉积,并伴随小胶质细胞激活;在6月龄时,AD模型组小鼠海马和PFC中神经递质水平持续降低,并伴随海马和PFC大量的Aβ沉积,海马神经元损伤以及胶质细胞激活水平的显著增加;同时出现认知行为障碍。结论 5×FAD小鼠在3月龄即出现部分神经递质水平紊乱,并随疾病进展进一步加剧,这一变化与Aβ沉积、神经炎症和神经元损伤密切相关。
Objective To investigate temporal relationships among neurotransmitter alterations,amyloid⁃beta(Aβ)deposition,and neuroinflammation in 5×FAD mice.Methods 5×FAD and C57BL/6J(WT)mice at 15 months(early pathological stage),3 months(mid⁃pathological stage),and 6 months(late pathological stage)were examined to establish a time⁃window model of Alzheimer’s disease(AD)progression.Each age and genotype group included 8 mice.Behavioral phenotyping was assessed using the Y⁃maze,novel object recognition,and open field tests.Immunofluorescence was used to detect Aβexpression and glial cell activation in the hippocampus and prefrontal cortex(PFC).HE staining was performed to observe hippocampal neuronal morphology.Neurotransmitter levels in the hippocampus and PFC were quantified via high⁃performance liquid chromatography.Results At 15 months,the hippocampal GABA level was lower in AD model group mice than in WT group mice.At 3 months,significant changes in several neurotransmitters,including acetylcholine,glutamate,serotonin,and 5⁃hydroxyindoleacetic acid(5⁃HIAA),were detected in the hippocampus of AD model group mice compared with WT group mice.At this stage,minor Aβdeposition and microglial activation were present in both the hippocampus and PFC.By 6 months,neurotransmitter levels further declined in the hippocampus and PFC of AD model group mice,accompanied by extensive Aβdeposition,pronounced hippocampal neuronal damage,and substantial glial cell activation.Cognitive impairments were also observed.Conclusions Neurotransmitter dysregulation in 5×FAD mice emerged by 3 months of age and worsened with disease progression.These alterations were closely associated with Aβdeposition,neuroinflammation,and neuronal injury.
作者
梁莹莹
李鑫
侯雪婷
刘冰琳
连丁丁
陈欢
侯宏卫
计敏
LIANG Yingying;LI Xin;HOU Xueting;LIU Binglin;LIAN Dingding;CHEN Huan;HOU Hongwei;JI Min(Hefei Institutes of Physical Science,Chinese Academy of Sciences,Hefei 230031,China;University of Science and Technology of China,Hefei 230026,China;Beijing Life Science Academy,Beijing 102299,China;China National Tobacco Quality Supervision&Test Center,Zhengzhou 450001,China;Key Laboratory of Tobacco Biological Effects,Zhengzhou 450001,China)
出处
《中国实验动物学报》
北大核心
2025年第11期1583-1597,共15页
Acta Laboratorium Animalis Scientia Sinica
基金
北京市自然科学基金-大兴创新联合基金重点研究专题项目(L246003)
北京生命科技研究院重点科技项目(2023000CB0030,2023100CB0060)。
