Screening Of Differentially Expressed TsRNA In Diabetic Foot Ulcers Wound And Bioinformatics Analysis

Objective:To detect the differential expression of tsRNA in diabetic foot ulcer wound and provide a theoretical basis for studying the pathogenesis of diabetic foot ulcers.Methods: Three cases of diabetic foot ulcer peri-incision tissues and normal control skin tissues were collected separately,and total RNA was extracted using Trizol method and then subjected to RNA highthroughput sequencing.Nine tsRNAs with significant differential changes were selected for real-time quantitative polymerase chain reaction(qRTPCR)validation.Among the validated tsRNAs,the three most significantly differentially expressed ones were screened for target gene prediction,gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)bioinformatics analysis.Results:1.193 differentially expressed(FC>1.5,P<0.05)tsRNAs were identified in diabetic foot ulcer wounds,of which 104 were up-regulated and 89 were down-regulated.2.The results of qRT-PCR validation of nine of the selected significantly different tsRNAs showed that seven validated tsRNAs were consistent with the sequencing data,including six up-regulated(tRF-LysCTT-001,tRF-Lys-CTT-002,tRF-Ala-AGC-008,tRF-Glu-CTC-003 tRFLys-CTT-005,tRF-Lys-CTT-008)and one down-regulated(tRF-SerGCT-008),and the remaining two tsRNAs(tRF-Gly-GCC-032,tRF-TrpCCA-005)were not statistically different.3.Bioinformatics analysis of the screened tRF-Ala-AGC-008,tRFLys-CTT-005 and tRF-Ser-GCT-008 may target 410,534 and 149 target genes,respectively.GO analysis and KEGG pathway analysis based on the predicted target genes showed that the differential tsRNAs were enriched to biological pathways focused on insulin resistance pathway,oxidative phosphorylation pathway,and cellular senescence pathway,which are involved in biological processes such as cell cycle,cell proliferation,cell migration and apoptosis.Conclusions: tsRNA was differentially expressed in diabetic foot ulcer wound tissue and normal control skin tissue,and the differentially expressed tsRNA may regulate diabetic foot ulcers formation and repair through target genes.