纹状体内注射AAV9-82Q诱导亨廷顿病小鼠模型构建及评价  

Construction and evaluation of Huntington′s disease mouse model induced by intrastriate injection of AAV9-82Q

作　　者：谭雅丽 黄林[2] 唐亚洲 王媛 罗永杰 TAN Ya-li;HUANG Lin;TANG Ya-zhou;WANG Yuan;LUO Yong-jie(University of Electronic Science and Technology of China,Chengdu 610054,China;Department of Neurology,Zigong First People′s Hospital,Zigong 643000,China;Department of Neurology,Jiangyou People′s Hospital,Jiangyou 621700,China;Department of Neurology,Sichuan Provincial People′s Hospital,Chengdu 610072,China)

机构地区：[1]电子科技大学,四川成都610054 [2]自贡市第一人民医院神经内科,四川自贡643000 [3]江油市人民医院神经内科,四川江油621700 [4]四川省医学科学院·四川省人民医院神经内科,四川成都610072

出　　处：《临床军医杂志》2024年第11期1145-1150,共6页Clinical Journal of Medical Officers

基　　金：四川省科技厅重点研发项目(2019YFS0214);自贡市科技局重点研发项目(2021YXY04)。

摘　　要：目的通过向纹状体内注射携带82个CAG重复序列的腺相关病毒(AAV)载体9型(AAV9-82Q)构建亨廷顿病(HD)小鼠模型并对其进行评价。方法将50只C57BL/6小鼠随机分入AAV9-82Q组(n=19,注射AAV9-82Q)、AAV9-GFP组(n=19,注射空白对照病毒AAV9-GFP)及正常组(n=12,不做任何处理)。造模后采用转棒实验和平衡木实验评估小鼠的运动功能,Y迷宫实验评估认知功能,免疫荧光染色检测纹状体mHtt和NeuN表达。结果从造模后4周开始,AAV9-82Q组小鼠从转棒上掉落的平均潜伏期短于正常组、AAV9-GFP组,差异有统计学意义(P<0.05)。在造模后18~24周,AAV9-82Q组小鼠通过平衡木的时间长于正常组、AAV9-GFP组,差异有统计学意义(P<0.05)。3组小鼠的自发交替转换率比较,差异无统计学意义(P>0.05)。AAV9-82Q组小鼠纹状体内可见明显的mHtt表达,但在正常组、AAV9-GFP组中未检测到mHtt;与正常组、AAV9-GFP组比较,AAV9-82Q组小鼠纹状体内NeuN表达未见明显减少。结论AAV9-82Q注射使小鼠出现运动障碍,纹状体过表达mHtt,成功构建HD小鼠模型。Objective To construct and evaluate a mouse model of Huntington′s disease(HD)by injecting adeno-associated virus(AAV)vector type 9 with 82 CAG repeats(AAV9-82Q)into the striatum.Methods Fifty C57BL/6 mice were randomly divided into AAV9-82Q group(n=19,injected with AAV9-82Q),AAV9-GFP group(n=19,injected with blank control virus AAV9-GFP)and normal group(n=12,without any treatment).After modeling,the motor function of mice was evaluated by rotating rod test and balance beam test,cognitive function was evaluated by Y maze test,and the expression of mHtt and NeuN in striatum was detected by immunofluorescence staining.Results Starting from 4 weeks after modeling,the average latency of mice falling from the rotating rod in AAV9-82Q group was shorter than that in normal group and AAV9-GFP group,and the difference was statistically significant(P<0.05).At 18-24 weeks after modeling,the time of passing balance beam in AAV9-82Q group was longer than that in normal group and AAV9-GFP group,and the difference was statistically significant(P<0.05).There was no significant difference in the spontaneous alternating conversion rate among the three groups(P>0.05).The expression of mHtt in striatum of AAV9-82Q group was obvious,but no mHtt was detected in normal group and AAV9-GFP group.Compared with normal group and AAV9-GFP group,the expression of NeuN in striatum of mice in AAV9-82Q group was not significantly reduced.Conclusion The injection of AAV9-82Q resulted in dyskinesia and overexpression of mHtt in the striatum,and the HD mouse model was successfully established.

关 键 词：亨廷顿病 腺相关病毒 亨廷顿基因 突变亨廷顿蛋白 载体 

分 类 号：R742[医药卫生—神经病学与精神病学]