Analysis Of Serum Differential MiRNAs And Their Target Genes In Patients With Arteriosclerosis

[Objective] Atherosclerosis(AS)is difficult to detect and diagnose in early stage.How to detect and prevent AS is a new research hotspot.In this study,by using the second generation sequencing technology,we obtained the miRNAs differential expression spectrum of AS and healthy human peripheral blood serum,from which we found out the miRNA molecules with significant differences;We studied the role of miRNAs differential expression in the early diagnosis of AS;and through the bioinformatics method,we analyzed the regulatory target genes,to clarify the role of serum miRNAs in the diagnosis of AS.[Method] Firstly,20 AS patients and 20 healthy adults’ peripheral blood samples were collected,in which the total RNA was extracted from the peripheral blood of three normal people and three patients with atherosclerosis randomly,and sent to the company to detect the miRNA differential expression level.The second generation sequencing technology was used to obtain the miRNA differential expression profile of AS patients’ serum.Three miRNA molecules were selected and verified by RT-qPCR.The target genes regulated by miRNA were analyzed and verified by bioinformatics method signal pathway analysis,go annotation,revealed its role in the occurrence and development of AS.[Result]:1.There was no significant difference in age and gender composition between as patients and healthy people(p> 0.05).The body mass index of AS patients was higher than that of normal people,which indicated that AS had the trend of obesity;The levels of total cholesterol,low density lipoprotein(LDL)and triglyceride in AS patients were higher than those in healthy people(p < 0.05),while the levels of high density lipoprotein(HDL)were lower than those in healthy people(p < 0.05).2.Obtain the miRNA differential expression profile:,We detected the miRNA differential expression profile between AS and healthy human serum,through the second generation sequencing.The results showed that there are 35 significant differences between the two,22 of which are up-regulated and 13 of which are down regulated.3.The results of real-time fluorescent quantitative PCR to verify the differential expression spectrum: select one up-regulatedmiR-221-3p and two down-regulated miR-126-5p and miR-143-3p to detect the expression in serum of AS patients and healthy adults.The results confirmed that the up-regulated trend of miRNA gene was consistent with the sequencing results.4.114 of common targeting genes regulated by miR-126-5p were detected by three software.106 signal pathways were involved and there were more than three genes involved in two KEGG signaling pathways.The Hippo Signal Pathway was closely related to AS.5.37 of common targeting genes regulated by miR-143-3p were detected by three softwares.35 signal pathways are involved and there were more than two genes are involved in five KEGG signal pathways.There are three pathways closely related to AS: glycerol phospholipid metabolism,MAPK signal pathway and Ras signal pathway.6.42 of common targeting target genes regulated by miR-221-3p were detected by three softwares.85 signal pathways are involved and there were more than three genes are involved in four KEGG signaling pathways.There are two pathways closely related to AS: nonalcoholic fatty liver and MAPK signaling pathway.[Conclusion]:1.The differential expression profile of miRNA related to atherosclerosis was obtained.2.Three miRNAs(miR-126-5p、 miR-143-3p、 miR-221-3p)may be potential markers of atherosclerosis.3.Three miRNAs(miR-126-5p、 miR-143-3p、 miR-221-3p)effected the occurrence and development of atherosclerosis through regulating those related signaling pathways.