阿尔茨海默病小鼠海马脑区PGC-1α阳性神经元活化模型的构建OA
Construction of a model of activated PGC-1α positive neurons in the hippocampus of Alzheimer's disease mice
目的:构建阿尔茨海默病(Alzheimer's disease,AD)小鼠海马脑区过氧化物酶体增殖物激活受体γ辅激活因子-1α(peroxisome proliferator-activated receptor-γ coactivator-1α,PGC-1α)活化的光遗传模型.方法:选择 24 只APP/PS1双转基因(AD模型)小鼠,依据病毒干预类型(AAV-Ppargc1α-hChR2/AAV-Control-hChR2)与光处理方式(光刺激/无光刺激)随机分为4组:AD+AAV-Control-hChR2+无光刺激组,AD+AAV-Control-hChR2+光刺激组,AD+AAV-Ppargc1α-hChR2+无光刺激组,AD+AAV-Ppargc1α-hChR2+光刺激组,每组6只.通过PCR鉴定,确定其基因型;通过脑区定点微灌注技术向AD小鼠海马齿状回区微灌注携带光敏蛋白hChR2的腺相关病毒,同时在注射位点植入光纤,病毒表达2周后,蓝光刺激,每5 min 1次,每次30 s,每天2 h,持续5 d.采用免疫荧光技术检测4组小鼠海马脑区Cfos+HA+细胞数,验证光敏蛋白的有效性;采用免疫荧光技术检测在相同光刺激条件下,AD小鼠海马脑区PGC-1α阳性细胞数.结果:PCR结果显示,同时扩增出APP及PS1条带的小鼠为AD双转基因小鼠.与无光刺激组(AD+AAV-Control-hChR2+无光刺激及AD+AAV-Pgc-1α-hChR2+无光刺激)相比,光刺激组(AD+AAV-Control-hChR2+光刺激及AD+AAV-Pgc-1α-hChR2+光刺激)小鼠海马脑区Cfos+HA+神经元数量增加(P＜0.01).在相同的光刺激条件下,与AAV-Control-hChR2+光刺激组相比,AAV-Pgc-1α-hChR2+光刺激组AD小鼠海马脑区PGC-1α阳性细胞数明显增加(t=9.613,P＜0.01).结论:成功构建AD小鼠海马脑区PGC-1α活化的光遗传模型.
Objective:To establish a optogenetic model of peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)activation in the hippocampal neurons of Alzheimer's disease(AD)mice.Methods:Twenty-four APP/PS1 double transgenic(AD model)mice were selected and randomly divided into 4 groups based on the type of viral intervention(AAV-Ppargc1α-hChR2/AAV-Control-hChR2)and light treatment(light stimulation/no light stimulation):AD+AAV-Control-hChR2+no light stimulation group,AD+AAV-Control-hChR2+light stimulation group,AD+AAV-Ppargc1α-hChR2+no light stimulation group,and AD+AAV-Ppargc1α-hChR2+light stimulation group,with 6 in each group.Genotypes were confirmed by PCR.Adeno-associated virus carrying the light-sensitive protein hChR2 was microinjected into the dentate gyrus of the hippocampus of AD mice,and optical fibers were implanted at the injection site.Two weeks after viral expression,blue light stimulation was applied at 5-minute intervals lasting for 30 seconds each,for 2 hours a day,for 5 days.Immunofluorescence was used to detect the number of Cfos+HA+cells in the hippocampus of the four groups of mice to verify the effectiveness of the light-sensitive protein;immunofluorescence was also used to detect the number of PGC-1α positive neurons in the hippocampus of AD mice under the same light stimulation conditions.Results:PCR results showed that mice with both APP and PS1 bands were AD double transgenic mice.Compared with the no light stimulation groups(AD+AAV-Control-hChR2+no light stimulation and AD+AAV-Pgc-1α-hChR2+no light stimulation),the number of Cfos+HA+neurons in the hippocampus of the light stimulation groups(AD+AAV-Control-hChR2+light stimulation and AD+AAV-Pgc-1α-hChR2+light stimulation)increased significantly(P＜0.01).Compared with AAV-Control-hChR2+light stimulation group,the expression of PGC-1α positive neurons in the hippocampus of AD mice in AAV-Pgc-1α-hChR2+light stimulation group was significantly increased(t=9.613,P＜0.01).Conclusion:A hippocampus-specific optogenetic model for PGC-1α activation in AD mice was successfully established.
王贻界;朱旭;康子涵;邹成智;王佳庆;潘雯;王佳;刘凤国
江苏大学医学院,江苏镇江 212013||江苏大学第四附属医院神经内科,江苏镇江 212001镇江市中医院疼痛科,江苏镇江 212000江苏大学医学院,江苏镇江 212013江苏大学医学院,江苏镇江 212013江苏大学医学院,江苏镇江 212013江苏大学医学院,江苏镇江 212013江苏大学医学院,江苏镇江 212013||江苏大学第四附属医院神经内科,江苏镇江 212001江苏大学第四附属医院神经内科,江苏镇江 212001
医药卫生
阿尔茨海默病光遗传学过氧化物酶体增殖物激活受体γ辅激活因子-1α海马
Alzheimer's diseaseoptogeneticsperoxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)hippocampus
《江苏大学学报（医学版）》 2026 (2)
100-104,5
江苏省中医药科技发展计划面上项目(MS2025107)江苏省检验医学重点实验室开放课题(JSKLM-Y-2025-15)
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