HGF修饰脂肪间充质干细胞和微囊化雪旺细胞移植对糖尿病创面血管新生的作用及机制研究

Therapeutic angiogenesis is the key strategy for the treatment of vascular occlusive diseases such as diabetic foot ulcers. The transplantation of vascular growth factor gene modified stem cells is an effective way to achieve this goal. Our studies have found that hepatocyte growth factor (HGF), as a specific growth factor of vascular endothelial cells (VEC), could induce adipose mesenchymal stem cells (Ad-MSCs) to form tube like structures and promote wound healing. It is reported that Schwann cells (SC) could secrete a variety of cytokines that promote axon growth and induce MSCs to differentiate into VEC. Accordingly, our team hypothesized that HGF gene modified Ad-MSCs combined with SC as a superimposed angiogenesis means could effectively promote DFU healing which characterized with vasculopathy and neuropathy. Based on previous studies, we aimed to explore the role and mechanism of HGF gene modified Ad-MSCs in directional differentiation and angiogenesis through protein detection, microcarrier culture and three-dimensional reconstruction etc. And establishing three-dimensional composite biomaterials combined with microencapsulation of SC, HGF gene modified Ad-MSCs and chitosan hydrogel to explore its effect on DFU angiogenesis and wound healing. So as to lay a theoretical foundation provide new ideas for clinical treatment of DFU.

治疗性血管新生是治疗糖尿病足溃疡（DFU）等血管闭塞性疾病的关键策略。移植血管生长因子基因修饰的干细胞是实现该策略的有效途径。项目组前期研究发，现肝细胞生长因子（HGF）作为血管内皮细胞（VEC）特异生长因子可诱导脂肪间充质干细胞（Ad-MSCs）形成管样结构，促进创面愈合。而文献报道，雪旺细胞（SC）可分泌多种细胞因子促进轴突生长并促使MSCs向VEC分化。据此课题组提出假设，HGF基因修饰的Ad-MSCs结合SC作为叠加性血管新生手段，将有效促进速血管和神经病变为主的DFU愈合。本项目拟在前期研究基础上，通过蛋白检测、微载体培养、三维重建等方法探讨HGF基因修饰的Ad-MSCs定向分化、促血管新生的作用及机制；并联合应用微囊化SC+HGF基因修饰的Ad-MSCs+壳聚糖水凝胶建立三维复合生物材料，探索其对DFU血管新生及创面愈合的作用。从而为临床治疗DFU奠定理论基础，提供新思路。

治疗性血管新生是治疗糖尿病足溃疡（DFU）等血管闭塞性疾病的关键策略。移植血管生长因子基因修饰的干细胞是实现该策略的有效途径。项目组前期研究发现肝细胞生长因子（HGF）可增强脂肪干细胞（ADSC）促血管新生的作用，外泌体在其中发挥着重要的通讯作用。施旺细胞（SC）可分泌多种细胞因子促进轴突生长改善糖尿病神经病变。课题组研究发现，ADSC外泌体可以促进创面成纤维细胞、角质形成细胞的迁移，血管内皮细胞的增殖及成管，从而加速创面愈合；HGF基因修饰的ADSC外泌体中HGF含量增加，具有更强的促进创面血管新生能力，能够有效改善糖尿病溃疡小鼠模型创面血供，促进愈合，该过程可能与激活PI3K/AKT信号通路，增加eNOS蛋白表达相关。在此基础上我们通过结合微囊化SC的创面局部神经营养作用，建联合应用微囊化SC+HGF基因修饰的ADSC+壳聚糖明胶水凝胶建立三维复合生物材料，证明其对DFU血管新生及创面愈合的作用。从而为临床治疗DFU奠定理论基础，提供新思路。