神经导向因子 1 可改善去神经支配的 2 型糖尿病小鼠脂肪干细胞的增殖、迁移和治疗效果。
Netrin-1 improves adipose-derived stem cell proliferation, migration, and treatment effect in type 2 diabetic mice with sciatic denervation.
机构信息
Department of Vascular Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
出版信息
Stem Cell Res Ther. 2018 Oct 25;9(1):285. doi: 10.1186/s13287-018-1020-0.
BACKGROUND
Diabetic peripheral neurovascular diseases (DPNVs) are complex, lacking effective treatment. Autologous/allogeneic transplantation of adipose-derived stem cells (ADSCs) is a promising strategy for DPNVs. Nonetheless, the transplanted ADSCs demonstrate unsatisfying viability, migration, adhesion, and differentiation in vivo, which reduce the treatment efficiency. Netrin-1 secreted as an axon guidance molecule and served as an angiogenic factor, demonstrating its ability in enhancing cell proliferation, migration, adhesion, and neovascularization.
METHODS
ADSCs acquired from adipose tissue were modified by Netrin-1 gene (NTN-1) using the adenovirus method (N-ADSCs) and proliferation, migration, adhesion, and apoptosis examined under high-glucose condition. The sciatic denervated mice (db/db) with type 2 diabetes mellitus (T2DM) were transplanted with N-ADSCs and treatment efficiency assessed based on the laser Doppler perfusion index, immunofluorescence, and histopathological assay. Also, the molecular mechanisms underlying Netrin-1-mediated proliferation, migration, adhesion, differentiation, proangiogenic capacity, and apoptosis of ADSCs were explored.
RESULTS
N-ADSCs improved the proliferation, migration, and adhesion and inhibited the apoptosis of ADSCs in vitro in the condition of high glucose. The N-ADSCs group demonstrated an elevated laser Doppler perfusion index in the ADSCs and control groups. N-ADSCs analyzed by immunofluorescence and histopathological staining demonstrated the distribution of the cells in the injected limb muscles, indicating chronic ischemia; capillaries and endothelium were formed by differentiation of N-ADSCs. The N-ADSCs group showed a significantly high density of the microvessels than the ADSCs group. The upregulation of AKT/PI3K/eNOS/P-38/NF-κB signaling pathways and secretion of multiple growth factors might explain the positive effects of Netrin-1 on ADSCs.
CONCLUSION
The overexpression of Netrin-1 in ADSCs improves proliferation, migration, and treatment effect in type 2 diabetic mice with sciatic denervation, which directs the clinical treatment of patients with DPNVs.
背景
糖尿病周围神经血管病变（DPNVs）是一种复杂的疾病，目前缺乏有效的治疗方法。脂肪来源的干细胞（ADSCs）的自体/同种异体移植是治疗 DPNVs 的一种有前途的策略。然而，移植的 ADSCs 在体内的活力、迁移、黏附和分化表现不佳，从而降低了治疗效果。轴突导向分子神经导向因子 1（Netrin-1）也具有血管生成因子的功能，能够增强细胞增殖、迁移、黏附和新生血管形成。
方法
采用腺病毒法对脂肪组织来源的 ADSCs 进行 Netrin-1 基因（NTN-1）修饰（N-ADSCs），并在高糖条件下检测细胞的增殖、迁移、黏附和凋亡情况。将 2 型糖尿病（db/db）模型的坐骨神经去神经小鼠移植 N-ADSCs，根据激光多普勒灌注指数、免疫荧光和组织病理学检测评估治疗效果。还探索了 Netrin-1 介导的 ADSCs 增殖、迁移、黏附、分化、促血管生成能力和凋亡的分子机制。
结果
N-ADSCs 可提高高糖条件下 ADSCs 的增殖、迁移和黏附能力，并抑制其凋亡。与 ADSCs 组和对照组相比，N-ADSCs 组的激光多普勒灌注指数升高。通过免疫荧光和组织病理学染色分析 N-ADSCs，结果显示细胞分布在注射肢体肌肉中，表明存在慢性缺血；N-ADSCs 分化形成了毛细血管和内皮细胞。N-ADSCs 组的微血管密度明显高于 ADSCs 组。AKT/PI3K/eNOS/P-38/NF-κB 信号通路的上调和多种生长因子的分泌可能解释了 Netrin-1 对 ADSCs 的积极作用。
结论
在坐骨神经去神经的 2 型糖尿病小鼠中，ADSCs 中转染 Netrin-1 可提高其增殖、迁移能力和治疗效果，为 DPNVs 患者的临床治疗提供了新的思路。
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