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

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 Δ E9 double transgenic mouse model of Alzhaimer's disease using real-time PCR. Methods Six-month-old APPswe/PS Δ 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 significance differences in the expressions of miRNA-135a-5p, miRNA-298-5p, 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 ΔE9 double transgenic mice, suggesting their important roles in the pathogenesis of Alzheimer disease.