Screening Of Differentially Expressed RNA And Possible CeRNA Mechanisms In Diabetic Foot By RNA-seq

Background:Diabetic foot is caused by diabetic patients with neuropathy and various degrees of peripheral vascular disease leading to lower limb infection,ulcer formation and / or deep tissue destruction.It has high morbidity,disability and mortality.However,the pathogenesis of diabetic foot is still unclear.With the continuous development of RNA-Seq technology,more and more pathogenesis of the diseases are gradually discovered.Sequencing technology has gradually become a hot spot.RNA-Seq technology can comprehensively and quickly obtain almost all transcripts including mRNA and ncRNA in a specific organ or tissue of a species.ncRNA does not encode proteins,but it can perform various biological functions at the level of RNA.ceRNA reveals a new mechanism for RNA-to-RNA interaction.In recent years,studies have also shown that ceRNA regulation mechanisms revealed new directions for disease pathogenesis and treatment.RNA-seq technology has achieved a series of results in the molecular etiology research of various diseases,but there are few related studies in diabetic foot.Therefore,it is expected that RNA-Seq technology will be used to analyze the transcriptome expression changes in peripheral blood of patients with diabetic foot and healthy people,in order to explore the underlying molecular mechanism of diabetic foot.Objective:RNA-Seq technology will be used to analyze the transcriptome expression changes in peripheral blood of patients with diabetic foot and healthy people,in order to find potential differential genes and ceRNA mechanisms and to identify the pathogenesis related to diabetic foot.Methods:（1）Peripheral blood samples from five diabetic foot patients with Wagner grades 2-3 were selected,including three males and two females.Five peripheral blood samples were selected from healthy people,including three males and two females.（2）RNA-Seq technology was used to sequence the entire transcriptome of diabetic foot patients and healthy people.（3）Analyzed the mRNA,miRNA,and lncRNA that were differentially expressed between the two transcripts to find potential differential genes and ceRNA mechanisms.（4）According to the GO and KEGG databases,the signal pathways involved in the differentially expressed genes were analyzed.Results:（1）In this study,3234 mRNAs,28 miRNAs and 2136 lncRNAs that were differentially expressed were found.（2）Differential expression data showed that NUTM2B-AS1 targets the NOTCH signaling pathway through sponge Hsa-miR-148a-3p.This may be a ceRNA regulatory mechanism in the pathogenesis of diabetic foot.（3）The sequencing results showed that MMP9 was highly expressed in patients with diabetic foot.（4）The miRNAs regulating MMP9 were Hsa-miR-4448 and Hsa-novel-159.The two miRNAs were differentially expressed in diabetic foot sequencing.（5）The lncRNAs regulating MMP9 were SLC12A5-AS1 and XLOC₀64360.The two lncRNAs were differentially expressed in diabetic foot sequencing.（6）The results of GO and KEGG functional analysis showed that diabetic foot may be closely related to MAPK signaling pathway,PI3K-AKT signaling pathway,cell adhesion molecules,neurotrophin signaling pathway,insulin resistance and RAS signaling pathway.Conclusion:Analyzed the transcriptome expression changes in peripheral blood of patients with diabetic foot and healthy people.Differential expression data showed that NUTM2B-AS1 targets the NOTCH signaling pathway through sponge Hsa-miR-148a-3p.This may be a ceRNA regulatory mechanism in the pathogenesis of diabetic foot.The miRNAs that regulate MMP9 were Hsa-miR-4448 and Hsa-novel-159,and the lncRNAs were SLC12A5-AS1 and XLOC₀64360.The two miRNAs and lncRNAs were differentially expressed in diabetic foot sequencing.