2026 年第 4 期 第 21 卷

Netrin-1过表达巨噬细胞来源外泌体促进糖尿病小鼠周围神经损伤后的血管新生和神经修复

Netrin-1 overexpressing macrophage-derived exosomes promote angiogenesis and nerve repair after peripheral nerve injury in diabetic mice

作者：王启明张欣怡黄群吴小雨刘俊超李维敏陆信武秦金保

英文作者：Wang Qiming Zhang Xinyi Huang Qun Wu Xiaoyu Liu Junchao Li Weimin Lu Xinwu Qin Jinbao

单位：上海交通大学医学院附属第九人民医院血管外科，上海200011

英文单位：Department of Vascular Surgery Shanghai Ninth People′s Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200011 China

关键词：周围神经损伤；外泌体；Netrin-1；血管新生

英文关键词：Peripheralnerveinjury；Exosomes；Netrin-1；Angiogenesis

• 摘要：

• 目的 探讨Netrin-1过表达巨噬细胞来源外泌体（N-Exos）对2型糖尿病（T2DM）小鼠坐骨神经损伤后的血管新生和神经修复的影响。方法 从小鼠下肢骨髓中分离骨髓来源巨噬细胞（BMDMs），使用腺病毒转染法使BMDMs过表达Netrin-1；检测过表达Netrin-1后巨噬细胞表型并提取其外泌体，通过透射电子显微镜、纳米颗粒追踪分析技术和蛋白质免疫印迹法对外泌体进行鉴定。将N-Exos与人脐静脉内皮细胞（HUVECs）体外高糖条件下共培养后检测N-Exos对HUVECs的增殖、凋亡、迁移能力、成管能力的影响；将18只8周龄雄性C57BL/KsJ-db/db小鼠完全随机分为3组：坐骨神经损伤组（对照组）、外泌体治疗组（Exos组）和Netrin-1过表达外泌体治疗组（N-Exos组）。构建T2DM小鼠坐骨神经损伤模型，评估N-Exos对T2DM小鼠坐骨神经损伤后神经修复的影响。结果 N-Exos组细胞增殖速率高于Exos组和对照组，N-Exos组HUVECs的24 h划痕愈合率高于Exos组和对照组，与N-Exos共培养的HUVECs小管形成相对累计长度长于Exos组和对照组(均P＜0.05)。TUNEL染色结果显示对照组HUVECs凋亡细胞数量最多，N-Exos组HUVECs凋亡细胞数量最少。T2DM小鼠体内实验证实，N-Exos组损伤坐骨神经内肿瘤坏死因子α相对荧光强度低于对照组和Exos组，髓鞘碱性蛋白和神经丝蛋白的相对荧光强度均高于对照组和Exos组，N-Exos组的损伤神经内CD31相对荧光强度高于对照组和Exos组。术后14 d，N-Exos组小鼠的腓肠肌萎缩指数显著低于Exos组和对照组［(15.7±1.0)%比(20.5±0.8)%、(25.7±2.5)%］(P=0.001)。结论 N-Exos可促进T2DM小鼠周围神经损伤后的血管新生和神经修复，为糖尿病患者周围神经损伤的临床治疗提供新的思路和理论依据。

• Objective To investigate the effect of Netrin-1 overexpressing macrophage-derived exosomes (N-Exos) on angiogenesis and nerve repair after sciatic nerve injury in type 2 diabetes mellitus (T2DM) mice. Methods Bone marrow-derived macrophages (BMDMs) were isolated from the lower limb bone marrow of mice, and adenovirus transfection was used to induce Netrin-1 overexpression in BMDMs. The phenotype of macrophages after Netrin-1 overexpression was detected, and their exosomes were extracted and identified by transmission electron microscopy, nanoparticle tracking analysis and Western blot. Human umbilical vein endothelial cells (HUVECs) were co-cultured with N-Exos under high-glucose conditions in vitro to detect the effects of N-Exos on the proliferation, apoptosis, migration and tube formation abilities of HUVECs. Eighteen 8-week-old male C57BL/KsJ-db/db mice were randomly divided into three groups: the sciatic nerve injury group (control group), the exosome treatment group (Exos group), and the Netrin-1 overexpressing exosome treatment group (N-Exos group). A sciatic nerve injury model of T2DM mice was established to evaluate the effect of N-Exos on nerve repair after sciatic nerve injury in T2DM mice. Results The cell proliferation rate in the N-Exos group was higher than that in the Exos group and the control group, the 24 h scratch healing rate of HUVECs in the N-Exos group was higher than that in the Exos group and the control group, and the relative cumulative length of tube formation of HUVECs co-cultured with N-Exos was longer than that in the Exos group and the control group (all P＜0.05). TUNEL staining results  showed that the number of apoptotic HUVECs was the largest in the control group and the smallest in the N-Exos group. In vivo experiments on T2DM mice confirmed that the relative fluorescence intensity of tumor necrosis factor-α in the injured sciatic nerve in the N-Exos group was lower than that in the control group and the Exos group, the relative fluorescence intensities of myelin basic protein and neurofilament protein were both higher than those in the control group and the Exos group, and the relative fluorescence intensity of CD31 in the injured nerve in the N-Exos group was higher than that in the control group and the Exos group. At 14 d after surgery, the gastrocnemius muscle atrophy index of mice in the N-Exos group was significantly lower than that in the Exos group and the control group ［(15.7±1.0)% vs (20.5±0.8)%, (25.7±2.5)%］(P=0.001).Conclusion N-Exos can promote angiogenesis and nerve repair after peripheral nerve injury in T2DM mice, which provides new ideas and a theoretical basis for the clinical treatment of peripheral nerve injury in diabetic patients.