Microarray-based MiRNAs Profiling In Lower Limb Arteriosclerosis Of Diabetes Rats

In clinical, we found that a growing number of diabetes patients with lower limb arteriosclerosis, clinical characteristics with lacunar claudication, rest pain, ischemic gangrene. So far, the detailed pathogenesis of atherosclerosis is not clarified, now more generally accepted "doctrine stress injuries" expressed its pathogenesis, but not involved in gene regulation. Due to the pathogenesis of blank in the field of gene regulation and intervention targets, it is impossible to determine the related genes affecting the effective prevention and treatment of diabetic lower limb arteriosclerosis.In recent years microRNAs become the most popular in the field of genetic research, about 30% of human genes is regulated by the microRNAs, a microRNA family may combine with up to 200 target genes, compared to a large number of target genes, regulating the role of microRNAs quantity is much less, so we can through microRNAs research to understand the pathogenesis of diabetic lower limb arteriosclerosis more easily, and it is also the great advantages of using microRNAs to investigate the gene expression regulation.Frist,this study is to ecplore the expression of microRNAs in diabetic lower limb arteriosclerosis,through the establishment of model rats, for subsequent exploring the pathogenesis of human suffering fiom diabetic lower limb arteriosclerosis lay animal model experimental basis.Objective:Through the establishment of model rats, microarray-based microRNAs profiling in the lower extremity arterial tissue between diabetic rat lower limb arteriosclerosis (DAS) and diabetic rat lower limb normal (DN), to explore the possible molecular mechanisms involved in aberrant microRNAs expression in DAS.Methods:The rats models of DAS and DN were successfully established. The respective lower extremity of the arterial vascular tissue was isolated. The total microRNAs were purified and had a hybridization detection by microRNA microarray. The results of chip scanning and data were analyzed and verified by RT-qPCR.Results:Ten microRNAs related to diabetic rats with lower limb arteriosclerosis 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, rno-miR-126a-3p were determined. All 10 abnormal expression of microRNAs were uupregulated. The validating results of RT-qPCR confirmed 9 of the microRNAs in line with chip expression, Just the rno-miR-335 showed opposite between PCR detection and microarray result. Chip shows that diabetes increases significantly lower limb arteriosclerosis rno-miR-206-3p, rno-miR-133a-5P, rno-miR-675-3p,99 times respectively,39 times,18 times, and P< 0.05, the expression of real-time quantitative also show a similar ratio. According to the data, it is worth making attention on these three microRNAs, as the key part of subsequent related function experiment of microRNAsConclusion:A group of microRNAs in diabetic rats suffering from lower limb arteriosclerosis plays an important role in the vascular atherosclerosis process, the abnormal expression of microRNAs is likely to affect the vascular atherosclerosis process.