Analysis Of Differential Expression Of Circular RNA In Human Retinal Vascular Endothelial Cells Cultured With High Glucose

Objective: The purpose of this study is to investigate the differential expression of circular RNAs(circ RNAs)in human retinal vascular endothelial cells(HRVECs) cultured in different glucose concentrations,and to explore the mechanism of circ RNAs in diabetic endothelial dysfunction.Methods: Human retinal vascular endothelial cells cultured in high glucose and normal glucose concentration for 24 hours were screened by circular RNA microarray.The differentially expressed circular RNA was screened out.Part of the differential expression circular RNA molecules were verified by q RT-PCR in cell samples of high glucose group,hypertonic control group and normal group.Finally,The miRNA molecule of the gene that possible downstream role of circular RNA was analyzed by bioinformatics.Results: Through circular RNA microarray analysis analysis,compared with HRVECs cultured in normal glucose concentration,448 circular RNA molecules were differentially expressed in HRVECs cultured with high sugar concentration(P<0.05,FC≥1.5),of which 182 circular RNAs were up-regulated,266 expressions were downregulated.Six of the most significant differentially expressed circular RNAs(hsa＿circ＿0002938,hsa＿circ＿0008036,hsa＿circ＿0001946,hsa＿circ＿0035277,hsa＿circ＿0008344,hsa＿circ＿0001874)were selected from among them for verification by q RT-PCR.The results showed that the relative expression levels of six kinds of circular RNAs among the three groups were significant Sexual difference(P<0.05);Compared with the normal group,there were no significant differences in the relative expression levels of each kind of circular RNAs in the hypertonic control group(P >0.05);compared with the normal group and the hypertonic control group,the expression levels of the three circular RNAs up-regulated in the microarray analysis were significantly increased in the high glucose group,and the difference was statistically significant(P<0.05).While the three circular RNAs down-regulated in the microarray analysis were in the high glucose group,the expression level was significantly down-regulated,and the difference was statistically significant(P<0.05).Among them,hsa＿circ＿0001946 was selected to analyze its downstream miRNA and construct a circ RNA-micro RNA network.The three miRNAs most likely to bind to hsa＿circ＿0001946 include hsa-miR-7-5p,hsa-miR-1246,and hsa-miR-139-3p.By exploring the role of hsa＿circ＿0001946 target miRNA in diseases,we speculate the possible potential regulatory and regulatory mechanisms of circular RNA in diabetes-related endothelial dysfunction.Conclusion: Circular RNA is differentially expressed in HRVECs cultured with high glucose,and may regulate the expression of target genes to participate in the occurrence and development of diabetic retinopathy,and play a regulatory role in endothelial dysfunction.