阿尔茨海默病病变相关脑区Pgc-1alpha敲除小鼠模型的构建  

Construction of Pgc-1alpha knockout mouse model in Alzheimer′s disease related pathological brain regions

作　　者：史厚珍 张维君 翟鸿儒 王贻界 王雨欣 刘子重[2] 王佳[1,2,3] SHI Houzhen;ZHANG Weijun;ZHAI Hongru;WANG Yijie;WANG Yuxin;LIU Zizhong;WANG Jia(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013;Department of Neurology,the Fourth Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212001;Zhenjiang Jieshengrui Biotechonology Co.,Ltd,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]江苏大学第四附属医院神经科,江苏镇江212001 [3]镇江杰胜瑞科技有限公司,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2024年第5期423-427,450,共6页Journal of Jiangsu University：Medicine Edition

基　　金：江苏省重点研发社会发展面上项目(BE2022782);镇江市政策引导类计划项目(GJ2021010);镇江“金山英才”高层次领军人才培养计划(第六期“169”工程)科研资助项目;镇江市重点研发产业前瞻与共性关键技术项目(SH2022019)。

摘　　要：目的:构建在阿尔茨海默病(Alzheimer′s disease,AD)病变相关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 receptorγcoactivator-1 alpha,Pgc-1alpha)敲除小鼠。方法:引入Pgc-1alpha条件性敲除杂合子小鼠(Pgc-1alphafl/+),通过自交获得Pgc-1alpha条件性敲除纯合子小鼠(Pgc-1alphafl/fl)。将Pgc-1alphafl/fl纯合子小鼠和在AD病变脑区(皮层、海马)特异性表达Cre重组酶的Cre工具鼠(Calb1-Cre)杂交,提取子代转基因小鼠基因组DNA,行PCR鉴定,确定其基因型;采用蛋白免疫印迹技术检测转基因小鼠与野生型小鼠脑区及肾脏PGC-1α蛋白表达。结果:双重PCR鉴定结果显示,同时扩增出400 bp及444 bp条带的小鼠为AD病变脑区Pgc-1alpha敲除纯合子小鼠(Pgc-1alphafl/fl::Calb1-Cre,Pgc-1alpha-/-)。蛋白免疫印迹结果显示,与野生型小鼠相比,条件性敲除小鼠脑区PGC-1α蛋白表达明显减少(P<0.05)。结论:成功构建在AD病变相关脑区Pgc-1alpha敲除小鼠。Objective:To constuct peroxisome proliferator-activated receptorγcoactivator-1 alpha(Pgc-1alpha)knockout mouse model in Alzheimer′s disease(AD)-related pathologic regions.Methods:Pgc-1alphafl/fl homozygous mice were obtained by crossing Pgc-1alphafl/+heterozygous offspring.Pgc-1alphafl/fl homozygous mice were hybridized with Cre tool mice(Calb1-Cre),which specifically expressed Cre recombinase in AD lesion brain regions(cortex and hippocampus),and the genetic DNA of progeny transgenic mice was extracted and identified by PCR.The expression of PGC-1αprotein in brain and kidney of transgenic mice and wild-type mice was detected by Western blotting.Results:Dual PCR analysis revealed that mice exhibiting simultaneous amplification of 400 bp and 444 bp bands were characterized by Pgc-1alpha deletion specific to AD-related brain regions(Pgc-1alphafl/fl::Calb1-Cre,Pgc-1alpha-/-).The results of Western blotting showed that the expression of PGC-1αprotein in the brain region of conditioned knockout mice was significantly decreased compared with that of wild-type mice(P<0.05).Conclusion:Conditional Pgc-1alpha knockout mice in AD-related pathological brain regions were successfully constructed.

关 键 词：过氧化物酶体增殖物激活受体γ辅激活因子-1α(Pgc-1alpha) 条件性敲除 小鼠 阿尔茨海默病 皮层 海马 

分 类 号：R749.16[医药卫生—神经病学与精神病学]