脂肪来源干细胞外泌体通过CCN2/PI3K/AKT途径促进糖尿病伤口愈合：治疗潜力与机制洞察

Adipose-derived stem cells extracellular vesicles enhance diabetic wound healing via CCN2/PI3K/AKT pathway: therapeutic potential and mechanistic insights.

作者信息

Zhou Yu-Lu, Ogura Shingo, Ma Hao, Liang Rong-Bin, Fang Shao-Yihan, Wang Yue-Ming, Wo Yan, Wang Wen-Jin, Liu De-Wu

机构信息

Medical Center of Burn Plastic and Wound Repair, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi, China.

Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

出版信息

Stem Cell Res Ther. 2025 Jun 15;16(1):304. doi: 10.1186/s13287-025-04354-x.

DOI:10.1186/s13287-025-04354-x

PMID:40518546

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12168405/

Abstract

BACKGROUND

Adipose-derived stem cells extracellular vesicles (ADSCs-EVs) hold significant promise in tissue repair and regeneration. While they have been reported to enhance diabetic wound healing, the precise mechanisms remain unclear.

METHODS

ADSCs-EVs were isolated via ultracentrifugation and characterized through transmission electron microscopy, Western blot, and nanoparticle tracking analysis. Their effects on human umbilical vein endothelial cells (HUVECs) and RAW 264.7 macrophages were assessed in vitro, focusing on cell proliferation, migration, tube formation, and macrophage polarization. A diabetic rat wound model was used to evaluate their therapeutic impact on wound healing and angiogenesis, with histological and immunofluorescence analyses. mRNA sequencing identified Cellular communication network factor 2(CCN2) as a key upregulated gene, leading to further exploration of its role in ADSCs-EVs-mediated angiogenesis and wound healing via the PI3K/AKT pathway. Gene silencing (si-CCN2) and pharmacological inhibition (LY294002) were employed both in vitro and in vivo.

RESULTS

ADSCs-EVs were successfully isolated and characterized. In vitro, ADSCs-EVs promoted HUVEC proliferation, migration, and tube formation, and facilitated macrophage polarization to the M2 phenotype. In vivo studies using a diabetic rat wound model confirmed the pro-healing effects of ADSCs-EVs, including enhanced angiogenesis, granulation tissue formation, and accelerated wound closure. mRNA sequencing revealed that CCN2 expression was significantly upregulated in diabetic wound tissues treated with ADSCs-EVs. Further experiments showed that inhibiting CCN2 expression (si-CCN2) or blocking the PI3K/AKT pathway (LY294002) partially suppressed HUVEC proliferation, migration, tube formation, and angiogenesis, and counteracted the pro-healing effects of ADSCs-EVs.

CONCLUSIONS

ADSCs-EVs promote diabetic wound healing through the CCN2/PI3K/AKT pathway, offering a promising therapeutic target for diabetic wound repair.

摘要

背景

脂肪来源干细胞外泌体（ADSCs-EVs）在组织修复和再生方面具有巨大潜力。虽然已有报道称其可促进糖尿病伤口愈合，但其确切机制仍不清楚。

方法

通过超速离心法分离ADSCs-EVs，并通过透射电子显微镜、蛋白质印迹法和纳米颗粒跟踪分析对其进行表征。在体外评估它们对人脐静脉内皮细胞（HUVECs）和RAW 264.7巨噬细胞的影响，重点关注细胞增殖、迁移、管腔形成和巨噬细胞极化。使用糖尿病大鼠伤口模型，通过组织学和免疫荧光分析评估它们对伤口愈合和血管生成的治疗作用。mRNA测序确定细胞通讯网络因子2（CCN2）为关键上调基因，进而通过PI3K/AKT途径进一步探索其在ADSCs-EVs介导的血管生成和伤口愈合中的作用。在体外和体内均采用基因沉默（si-CCN2）和药物抑制（LY294002）。

结果

成功分离并表征了ADSCs-EVs。在体外，ADSCs-EVs促进HUVEC增殖、迁移和管腔形成，并促进巨噬细胞极化为M2表型。使用糖尿病大鼠伤口模型的体内研究证实了ADSCs-EVs的促愈合作用，包括增强血管生成、肉芽组织形成和加速伤口闭合。mRNA测序显示，在接受ADSCs-EVs治疗的糖尿病伤口组织中CCN2表达显著上调。进一步实验表明，抑制CCN2表达（si-CCN2）或阻断PI3K/AKT途径（LY294002）可部分抑制HUVEC增殖、迁移、管腔形成和血管生成，并抵消ADSCs-EVs的促愈合作用。