Application Of MicroRNA Analysis Based On High-throughput Sequencing Technology In Biomarkers Of HFpEF Patients

Objective:In this study,miRNA differentially expressed in peripheral blood between patients with heart failure with preserved ejection fraction(HFpEF)and healthy individuals were analyzed.To explore miRNA related to HFpEF,predict their target genes and conduct pathway enrichment analysis,and screen out miRNA with potential for diagnosis of HFpEF.Methods:1.A total of 30 HFpEF patients hospitalized in the Department of general Practice and cardiovascular Medicine of our hospital from January 2021 to December 2022 and 30 healthy controls who underwent physical examination in the physical examination center during the same period were selected.Clinical data of all the study subjects were collected and compared.Ten patients and five healthy controls were selected to construct a small RNA high-throughput library,and the differential expression of miRNA(DEMs)between the HFpEF patients and the healthy controls were analyzed.Target gene prediction was performed for the DEMs using the miRBase database.HFpEF related genes were obtained by comparison between Genecards database and Online Mendelian Inheritance in Man(OMIM)database,and core target genes were obtained by associating with predicted target genes.GO and KEGG enrichment analysis and annotation of core target genes were performed using gene ontology(GO)database and Matesacpe database,respectively,and R language was used to map miRNA-m RNA interaction related to the regulation of HFpEF.2.During the q RT-PCR validation phase,3 of the remaining 20 patients and 3 of the 25 healthy individuals were randomly selected from the group,respectively,to observe the comparative expression trend of DEMs in the HFpEF group and the control group.Results :1.43 DEMs were obtained by small RNA sequencing analysis,of which 4 were up-regulated and 39 were down-regulated in the HFpEF group.A total of 15034 target genes were predicted,including 30 core target genes related to HFpEF and 41 corresponding core miRNA.KEGG pathway enrichment analysis of 30 core target genes showed that,These genes are mainly concentrated in thiamine metabolism,thermogenesis,terpenoid skeleton biosynthesis,linolenic acid metabolism,fatty acid chain elongation system,linoleic acid metabolism,pentose acid pathway,breast cancer,neurodegenerative pathes-multiple diseases,PD-L1 expression in cancer and PD-1 checkpoint pathway,aldosterone regulation of sodium reabsorption,Staphylococcus aureus infection,Parkinson’s disease,JAK-STAT signaling pathway,hepatocellular carcinoma,fat digestion and absorption,American trypanosomiasis pathway.2.The core DEMs m RNA regulation map was established,and the expression levels of has-miR-3130-3p and has-miR-208b-3p in peripheral circulating blood of HFpEF patients were higher than those of healthy controls.The expression levels of 39 core DEMs,including has-miR-144-3p,has-miR-190-5p,and has-miR-374a-3p,decreased compared with the healthy control group.3.In the q RT-PCR validation phase,3 out of 20 HFpEF patients and 3out of 25 healthy controls were randomly selected for validation,to observe the expression trend of the top 7 DEMs in terms of abundance in the HFpEF group and the control group.Conclusion:1.There were 41 differentially expressed core DEMs with differential expression of free miRNA in peripheral blood between patients with HFpEF and healthy individuals,of which 2 core DEMs were up regulated in the HFpEF group,thirty-nine core DEMs were down regulated in HFpEF group.2.Up-regulated has-miR-3130-3p and down-regulated has-miR-144-3p and has-miR-190-5p in HFpEF are key miRNAs,which have the diagnostic potential of HFpEF.