实时荧光定量PCR检测转基因阿尔茨海默病小鼠脑组织中miRNAs的差异表达  

Real-time PCR for detecting differential expressions of microRNAs in the brain of a transgenic mouse model of Alzheimer's disease

作　　者：田密[1] 丁宇[1] 侯德仁[1] 邓炎尧[1] 李维[1] 奉夏露[1] 

机构地区：[1]中南大学湘雅三医院神经内科,湖南长沙410013

出　　处：《南方医科大学学报》2013年第2期262-266,共5页Journal of Southern Medical University

基　　金：湖南省科技厅社会发展支撑计划课题(009SK3175;2012SK3218)

摘　　要：目的通过实时荧光定量PCR方法检测APPswe/PS△E9双转基因小鼠和对照小鼠脑组织中miRNA-135a-5p、miRNA-135a-2-3p、miRNA-298-5p、miRNA-466b-3p和miR-669f-3p的表达。方法选取6月龄大小的APPswe/PS△E9双转基因小鼠为实验组,同月龄同种系的野生型小鼠C57为对照组,分别提取其脑组织总miRNA,应用实时荧光定量PCR方法检测两组小鼠脑组织中miRNA-135a-5p、miRNA-135a-2-3p、miRNA-298-5p、miRNA-466b-3p和miR-669f-3p的表达。结果上述5种miRNAs在实验组与对照组小鼠中的相对表达量分别为0.73±0.27、1.08±0.58,2.47±6.15、1.65±0.67,0.72±0.14、1.31±0.73,0.57±0.34、1.06±0.35,0.63±0.26、1.02±0.18。其中有统计学意义的是miRNA-135a-5p、miRNA-298-5p、miRNA-466b-3p和miR-669f-3p(P≤0.05)。结论 miRNA-135a-5p、miRNA-298-5p、miRNA-466b-3p和miR-669f-3p在APPswe/PS△E9双转基因小鼠中有差异表达,可能在阿尔茨海默病的发病过程中起着重要作用。Objective To detect the expression of miRNA-135a-5p, miRNA-135a-2-3p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p in the brain tissue of the APPswe/PS △ E9 double transgenic mouse model of Alzhaimer＇s disease using real-time PCR. Methods Six-month-old APPswe/PS △ E9 double transgenic mice and wild-type C57 mice of the same species were examined for the expressions of miRNA-135a-5p, miRNA-135a-2-3p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p in the brain tissue using real-time PCR. Results The relative expression levels of the 5 miRNAs in the transgenic versus the wild-type mice were 0.73±0.27 vs 1.08±0.58, 2.47±6.15 vs 1.65±0.67, 0.72±0.14 vs 1.31±0.73, 0.57±0.34 vs 1.06±0.35, and 0.63＋0.26 vs 1.02±0.18, respectively,showing sign/ficance differences in the expressions of miRNA-135a-Sp, miRNA-298-Sp, miRNA-466b-3p, and miR-669f-3p between the two groups （P〈0.05）. Conclusion miRNA-135a-5p, miRNA-298-5p, miRNA-466b-3p and miR-669f-3p are expressed differentially in APPswe/PS △ E9 double transgenic mice, suggesting their important roles in the pathogenesis of Alzheimer disease.

关 键 词：阿尔茨海默病 转基因阿尔茨海默病小鼠 miRNA 实时荧光定量PCR 

分 类 号：R749.16[医药卫生—神经病学与精神病学]