Gu Yuan, Mu Zelan, Chen Yuanzheng, Wu Can, Shi Jie, Bai Nan
School of Clinical Medicine, Shandong Second Medical University, Weifang, China.
Department of Burns and Plastic Surgery, Emergency General Hospital, Beijing, China.
Front Cell Dev Biol. 2024 Sep 13;12:1468220. doi: 10.3389/fcell.2024.1468220. eCollection 2024.
Diabetes mellitus (DM), a chronic metabolic disease characterized by elevated blood sugar, leads to delayed or non-healing wounds, increasing amputation risks, and placing a significant burden on patients and society. While extensive research has been conducted on adipose-derived stem cells (ADSCs) for promoting wound healing, there is a scarcity of studies focusing on diabetic wounds, particularly those employing proteomics and bioinformatics approaches.
This study aimed to investigate the mechanisms by which ADSCs promote diabetic wound healing using proteomics and bioinformatics techniques.
Healthy rat fat tissue was used to isolate ADSCs. A T2DM rat model with back wounds was established. The experimental group received ADSC injections around the wound, while the control group received PBS injections. Wound healing rates were documented and photographed on days 0, 3, 7, 10, and 14. On day 7, wound tissues were excised for HE and Masson's staining. Additionally, on day 7, tissues were analyzed for protein quantification using 4D-DIA, with subsequent GO and KEGG analyses for differentially expressed proteins (DEPs) and protein-protein interaction (PPI) network analysis using STRING database (String v11.5). Finally, Western blot experiments were performed on day 7 wounds to verify target proteins.
In all measured days postoperatively, the wound healing rate was significantly higher in the ADSC group than in the PBS group (day 7: < 0.001, day 10: = 0.001, day 14: < 0.01), except on day 3 ( > 0.05). Proteomic analysis identified 474 differentially expressed proteins, with 224 key proteins after PPI analysis (78 upregulated and 146 downregulated in the ADSC group). The main cellular locations of these proteins were "cellular anatomical entity" and "protein-containing complex", while the biological processes were "cellular processes" and "biological regulation". The primary molecular functions were "binding" and "catalytic activity", with GO enrichment focused on "Wnt-protein binding", "neural development", and "collagen-containing extracellular matrix". Further analysis of PPI network nodes using LASSO regression identified Thy1 and Wls proteins, significantly upregulated in the ADSC group, as potentially crucial targets for ADSC application in diabetic wound treatment.
糖尿病（DM）是一种以血糖升高为特征的慢性代谢性疾病，可导致伤口愈合延迟或不愈合，增加截肢风险，给患者和社会带来沉重负担。虽然针对脂肪来源干细胞（ADSCs）促进伤口愈合已开展了大量研究，但针对糖尿病伤口的研究较少，尤其是采用蛋白质组学和生物信息学方法的研究。
本研究旨在运用蛋白质组学和生物信息学技术探究ADSCs促进糖尿病伤口愈合的机制。
采用健康大鼠脂肪组织分离ADSCs。建立背部有伤口的T2DM大鼠模型。实验组在伤口周围注射ADSCs，对照组注射PBS。在第0、3、7、10和14天记录伤口愈合率并拍照。在第7天，切除伤口组织进行HE和Masson染色。此外，在第7天，使用4D-DIA对组织进行蛋白质定量分析，随后对差异表达蛋白（DEPs）进行GO和KEGG分析，并使用STRING数据库（String v11.5）进行蛋白质-蛋白质相互作用（PPI）网络分析。最后，在第7天的伤口上进行蛋白质印迹实验以验证靶蛋白。
术后所有测量天数中，除第3天外（P>0.05），ADSC组的伤口愈合率均显著高于PBS组（第7天：P<0.001，第10天：P = 0.001，第14天：P<0.01）。蛋白质组学分析鉴定出474个差异表达蛋白，经PPI分析后有224个关键蛋白（ADSC组中78个上调，146个下调）。这些蛋白的主要细胞定位是“细胞解剖实体”和“含蛋白质复合物”，而生物学过程是“细胞过程”和“生物调节”。主要分子功能是“结合”和“催化活性”，GO富集集中在“Wnt蛋白结合”、“神经发育”和“含胶原蛋白的细胞外基质”。使用LASSO回归对PPI网络节点进行进一步分析，确定在ADSC组中显著上调的Thy1和Wls蛋白是ADSCs应用于糖尿病伤口治疗的潜在关键靶点。
