基于ceRNA网络的糖尿病足溃疡关键miRNA筛选及功能验证OA

Background and Aims:Impaired wound healing in diabetic foot ulcers(DFUs)is a major cause of amputation and mortality in affected patients,and its underlying molecular regulatory mechanisms remain incompletely understood.This study aimed to identify key microRNAs(miRNAs)associated with DFU healing through the integration of a competing endogenous RNA(ceRNA)network and weighted gene co-expression network analysis(WGCNA),and to investigate their biological functions in DFU. Methods:Expression profile datasets of circRNAs,lncRNAs,miRNAs,and mRNAs related to DFU were obtained from the Gene Expression Omnibus(GEO)database.Differentially expressed RNAs were identified using the limma package,and a ceRNA regulatory network was constructed.WGCNA was subsequently performed to identify key modules and hub genes associated with DFU healing,followed by the establishment of a hub ceRNA subnetwork.Quantitative real-time PCR(qRT-PCR)was used to determine the expression levels of candidate miRNAs in DFU and control tissues.An in vitro high-glucose model was established using human umbilical vein endothelial cells(HUVECs).Following transfection with miRNA mimics or inhibitors,the effects of hsa-miR-155-5p on cell proliferation,migration,invasion,and angiogenesis were evaluated.Western blot analysis was performed to detect the expression of proteins involved in the PI3K/Akt signaling pathway. Results:A total of 66 differentially expressed miRNAs were identified,and a DFU-related ceRNA regulatory network was constructed.WGCNA identified a key module significantly associated with DFU healing and six hub genes,namely PDE9A,PADI2,PRR15L,B3GALT5,TMPRSS2,and NWD1.Integration of the ceRNA network further identified hsa-miR-155-5p,hsa-miR-204-5p,and hsa-miR-302d-3p as candidate miRNAs.qRT-PCR validation demonstrated that hsa-miR-155-5p expression was significantly lower in DFU tissues than in control tissues(P<0.05).In vitro experiments showed that overexpression of hsa-miR-155-5p significantly enhanced the proliferation,migration,invasion,and tube formation abilities of HUVECs under high-glucose conditions,while simultaneously increasing the expression levels of phosphorylated PI3K and Akt proteins(both P<0.05). Conclusion:Integrated analysis based on the ceRNA network and WGCNA identified hsa-miR-155-5p as a key miRNA associated with DFU healing.hsa-miR-155-5p may promote endothelial cell function and angiogenesis through activation of the PI3K/Akt signaling pathway,suggesting its potential as a therapeutic target for DFU.