Correlation Study Between MiRNA Expression Profiles In Platelet-derived Exosomes And Coronary In-stent Restenosis

Objective: Small RNA gene sequencing was used to determine the difference in the expression of platelet-derived exosomes’(PDEs)microRNA(miRNA)between patients with in-stent restenosis(ISR)and patients without ISR,and to evaluate the significance of differential expression of miRNAs in PDEs as the diagnostic biomarkers of coronary artery ISR.Method: According to the inclusion and exclusion criteria of the experimental design,26 patients from the Department of Cardiology,Zhongda Hospital Affiliated to Southeast University School of Medicine from January 2020 to January 2021 were selected.The patient’s blood was collected to isolate platelets.Then PDEs,were separated by differential centrifugation and verified.Next Illumina HiSeq2000 sequencing was used to analyze the expression profile of miRNAs in PDEs of 3random patients with coronary artery ISR and 3 patients without coronary artery ISR.The difference of miRNAs and the prediction and enrichment of miRNAs target genes between the two groups were analyzed.The differential expression of miRNAs in PDEs of 20 patients was verified by qRT-PCR.And CCK-8 and Transwell chamber were used to detect the effects of miRNA-133A-3p mimics on the proliferation and migration of vascular smooth muscle cells(VSMC).Finally,the potential function of differentially expressed miRNAs after verification is predicted by using miRNA-gene-Go network and miRNA-gene-KEGG network. Result: The results showed that 345 kinds of miRNAs were up-regulated and 584 kinds of miRNAs were down-regulated.According to the analysis of differential expression,there were significant differences in 5 miRNAs(p<0.05).Among them,one kind of miRNA expression was significantly up-regulated while four were significantly down-regulated.After verification by qRT-PCR,the expression of miRNA PC-3p-75179＿24 was up-regulated and the expression of miRNA-133a-3p＿R+1 and miRNA-5p＿R-1 was down-regulated.Mi RNA-133A-3p mimics could significantly inhibit the proliferation and migration of VSMC.The results also show three validated network diagrams of the interaction between miRNA-gene-GO and miRNA-gene-KEGG,and the potential functions of these differentially expressed miRNAs are predicted and discussed.Conclusion: This study found that the expression of miRNAs in PDEs of patients with coronary artery ISR changed,in which the expression of miRNA PC-3p-75179＿24 was specifically up-regulated,while the expression of miRNA-133a-3p＿R+1 and miRNA-589-5p＿R-1 were down-regulated.And miRNA-133A-3p mimics could significantly inhibit the proliferation and migration of VSMC.It supports the need to explore the potential of these miRNAs as biomarkers and the pathological effects in ISR.