应用微阵列芯片分析糖尿病大鼠下肢动脉硬化miRNAs差异表达谱

Microarray-based miRNAs profiling in lower limb arteriosclerosis of diabetic rats

目的:通过建立糖尿病大鼠模型,检测糖尿病大鼠动脉硬化下肢与糖尿病大鼠无动脉硬化下肢的动脉组织中微小RNAs(miRNAs)的差异表达情况,探究异常表达的miRNAs参与糖尿病大鼠下肢动脉硬化可能的分子机制。方法:选取建模成功的患有糖尿病下肢动脉硬化和患有糖尿病但无下肢动脉硬化的大鼠,取出下肢动脉组织,分别提取总的miRNAs,用miRNAs微阵列芯片进行杂交检测,经过芯片扫描和数据分析,再用RT-q PCR验证芯片的扫描分析结果,最终获得糖尿病大鼠下肢动脉硬化的miRNAs差异表达谱。结果:筛选出10个与糖尿病大鼠下肢动脉硬化有关的miRNAs,即rno-miR-206-3p、rno-miR-133a-5p、rno-miR-133b-3p、rno-miR-133a-3p、rno-miR-325-5p、rno-miR-675-3p、rno-miR-411-5p、rno-miR-329-3p、rno-miR-335和rno-miR-126a-3p,这10个异常表达的miRNAs都上调。RT-q PCR证实,其中9个miRNAs与芯片检查结果一致,仅仅rno-miR-335的PCR检测结果与基因芯片检测结果相反,表达下调。结论:有一群miRNAs在糖尿病大鼠下肢动脉硬化中起着重要作用,其表达过程很可能影响着动脉硬化的进程。

AIM： To establish the profiling of microRNAs（ miRNAs） in the lower extremity arterial tissue between diabetic rats with lower limb arteriosclerosis（ DAS） and diabetic rats with normal lower limb（ DN）,and to explore the possible molecular mechanisms involved in aberrant miRNA expression in DAS. METHODS： The rat models of DAS and DN were successfully established. The respective lower extremity arterial tissue was isolated. The total miRNAs were purified for a hybridization detection by miRNA microarray. The results of chip scanning and data were analyzed and verified by RT-q PCR. RESULTS： Ten miRNAs related to DAS,including rno-miR-206-3p,rno-miR-133a-5p,rno-miR-133b-3p,rno-miR-133a-3p,rno-miR-325-5p,rno-miR-675-3p,rno-miR-411-5p,rno-miR-329-3p,rno-miR-335 and rnomiR-126a-3p,were determined. All 10 abnormally expressed miRNAs were up-regulated. The validating results of RTq PCR confirmed 9 of the miRNAs in line with chip expression. Just rno-miR-335 showed the opposite between PCR detection and microarray result. CONCLUSION： A group of miRNAs in diabetic rats suffering from lower limb arteriosclerosis plays an important role in the vascular atherosclerosis process. The abnormal expression of miRNAs is likely to affect the vascular atherosclerosis process.