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Updated: Aug 19, 2025
Protocol to Create Chronic Wounds in Diabetic Mice
Published on: September 25, 2019
Screening and analysis of differentially expressed circRNAs and miRNAs in chronic diabetic extremity wounds
Xiaoliang Li1,2, Dan Lv1, Jiangfan Xie2
1Medical Center of Burn plastic and wound repair, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Abstract:
Increasing studies have shown that circular RNAs (circRNAs) and microRNAs (miRNAs) are related to the development of endocrine and metabolic diseases. However, there are few reports on the expression of circRNAs and miRNAs and their related co-expression and the expression of competitive endogenous RNA (ceRNA) in diabetic chronic refractory wounds. In this study, we compared the differential expression of circRNAs and miRNAs in diabetes chronic refractory wounds and normal skin tissues by high-throughput gene sequencing, and screened the differentially expressed circRNAs and miRNAs. Five abnormally expressed circRNAs and seven abnormally expressed miRNAs were detected by reverse transcription quantitative polymerase chain reaction PCR (RT-qPCR)to verify the results of RNA sequencing. We applied gene ontology (GO) to enrich and analyze dysregulated genes and elucidated their main functions via the Kyoto encyclopedia of genes and genomes analysis (KEGG). We constructed coding noncoding gene co-expression networks and ceRNA networks based on significantly abnormally expressed genes. According to the results of coding noncoding gene co-expression network analysis, hsa_circRNA_104175, hsa_circRNA_ 001588, hsa_circRNA_104330, hsa_circRNA_ 100141, hsa_circRNA_103107, and hsa_ circRNA_102044 may be involved in the regulation of the chronic intractable wound healing process in diabetes mellitus. This is particularly true in the regulation of vascular smooth muscle contraction-related pathways and the actin cytoskeleton, which affect the healing of chronic intractable wounds in diabetes. MiR-223-5p, miR-514a-3p, miR-205-5p, and miR-203-3p, which each have a targeting relationship with the above circRNAs, regulate the metabolism of nitrogen compounds in wound tissue by regulating NOD-like receptor signaling pathways, signaling pathways regulating the pluripotency of stem cells, microRNAs in cancer, and ECM-receptor interaction. This study showed circRNAs, miRNAs, and their network are associated with the development of chronic intractable wounds in diabetes, and our research identified the goals for new molecular biomarkers and gene therapy.
Insights
This study reveals circular RNAs (circRNAs) and microRNAs (miRNAs) are key players in diabetic chronic refractory wound healing. Their complex interactions influence pathways critical for wound repair, offering new therapeutic targets.
Area of Science:
- Molecular Biology
- Genetics
- Endocrinology
Background:
- Circular RNAs (circRNAs) and microRNAs (miRNAs) are implicated in endocrine and metabolic diseases.
- Limited understanding exists regarding circRNA and miRNA expression and their competitive endogenous RNA (ceRNA) networks in diabetic chronic refractory wounds.
Purpose of the Study:
- To investigate the differential expression of circRNAs and miRNAs in diabetic chronic refractory wounds.
- To construct co-expression and ceRNA networks to elucidate their roles in wound healing.
- To identify potential molecular biomarkers and therapeutic targets for diabetic wound management.
Main Methods:
- High-throughput gene sequencing to identify differentially expressed circRNAs and miRNAs.
- Reverse transcription quantitative polymerase chain reaction (RT-qPCR) for validation.
- Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses for functional enrichment.
- Construction of coding-noncoding gene co-expression and ceRNA networks.
Main Results:
- Five circRNAs and seven miRNAs showed abnormal expression in diabetic chronic refractory wounds.
- Identified circRNAs (e.g., hsa_circRNA_104175) and miRNAs (e.g., miR-223-5p) potentially regulate vascular smooth muscle contraction, actin cytoskeleton, and nitrogen compound metabolism.
- Constructed co-expression and ceRNA networks highlight the intricate regulatory mechanisms involved in diabetic wound healing.
Conclusions:
- CircRNAs, miRNAs, and their associated networks play a significant role in the pathogenesis of chronic refractory wounds in diabetes.
- The identified circRNAs and miRNAs represent promising molecular biomarkers for diagnosis and potential targets for gene therapy in diabetic wound healing.
