内皮祖细胞成血管及神经营养双重效应对糖尿病周围神经病变的影响及黄芪组分干预研究

Diabetic Peripheral Neuropathy（DPN）is characterized by deficiency of angiogenic factors and neurotrophic factors,Endothelial Progenitor Cells (EPCs) can be an optimal candidate for treating DPN because they possess paracrine properties that encompass both angiogenic and neurotrophic effects.The subject to preparation of bone marrow-derived EPCs,Build diabetic rat model,Local transplantation of EPCs after The Astragalus component intervention cultured,Measurements of nerve conduction velocity and tail-flick temperatures;Perfusion of sciatic vasa nervorum was measured with a laser Doppler perfusion imager(LDPI)system;assessed the neurotrophic vascular functional changes; The neurotrophic vascular functional changes was assessed by The Fluorescent imaging of blood vessels in sciatic nerves and femoral muscles; Studied the engraftment and transdifferentiation characteristics of EPCs by Flow cytometry and immunohistochemistry; The impact of EPCs on endothelial cell and Schwann cell proliferation and apoptosis was detected by Immunohistochemistry, cell proliferation/apoptosis kit;The levels of angiogenic and neurotrophic factors expression was detected by Real-time RT-PCR, Western blot.This project was designed to observe the Dual Angiogenic and Neurotrophic Effects of Endothelial Progenitor Cells local transplantation on Diabetic Peripheral Neuropathy and its Mechanisms of Astragalus Membranaceus intervention and provide a theoretical basis for the practical application of Astragalus.

糖尿病周围神经病变（DPN）以缺少成血管因子和神经营养因子为特征，内皮祖细胞（EPCs）具备成血管和神经营养双重效应，其局部移植治疗对于DPN的防治将提供良好的前景。本课题制备骨髓来源EPCs；构建糖尿病大鼠模型，黄芪组分干预培养后EPCs局部移植，检测坐骨神经传导速度，甩尾温度阈值；激光多普勒灌注影像系统评估坐骨神经滋养血管血流灌注；坐骨神经血管荧光显像评估神经滋养血管功能变化；流式细胞术、免疫组织化学检测EPCs移植特性及转分化特性；免疫组织化学、细胞增殖/凋亡试剂盒检测EPCs对内皮细胞和雪旺细胞增殖、凋亡的影响；Real-time RT-PCR、Western Blot检测成血管因子及神经营养因子水平表达。通过本课题研究，旨在观察EPCs局部移植治疗DN的成血管及神经营养双重效应及黄芪组分干预对其影响，为黄芪治疗DPN的实际应用提供理论依据，为DPN防治提供新的治疗思路和手段。

糖尿病周围神经病变（DPN）以缺少成血管因子和神经营养因子为特征，内皮祖细胞（EPCs）具备成血管和神经营养双重效应，其局部移植治疗对于DPN的防治将提供良好的前景。本课题制备骨髓来源EPCs；构建糖尿病大鼠模型，黄芪组分干预培养后EPCs局部移植，检测坐骨神经传导速度，甩尾温度阈值；细胞增殖试剂盒检测EPCs对内皮细胞和雪旺细胞增殖的影响；RT-PCR、免疫组化检测成血管因子及神经营养因子水平表达。通过本课题研究，旨在观察EPCs局部移植治疗DN的成血管及神经营养双重效应及黄芪组分干预对其影响，为黄芪治疗DPN的实际应用提供理论依据，为DPN防治提供新的治疗思路和手段。.研究表明：APS体外干预大鼠骨髓源性EPCs，能够影响其数量、增殖、黏附、分化、及周期的变化；APS体外体外干预大鼠骨髓源性EPCs共培养能够有效促进SCs、HUVEC的增殖；APS体外干预大鼠骨髓源性EPCs局部移植能够促进DPN大鼠模型坐骨神经传导速度及甩尾温度阈值的变化；APS体外干预大鼠骨髓源性EPCs局部移植能够促进DPN大鼠模型坐骨神经成血管因子及神经营养因子水平表达；APS体外干预大鼠骨髓源性EPCs局部移植能够促进SCs、HUVEC的增殖和延缓其凋亡。.通过本项目的系统研究，目前已基本阐明糖大鼠骨髓源性EPCs与SCs、HUVEC共培养能够有效促进细胞增殖；大鼠骨髓源性EPCs局部移植能够有效改善DN大鼠模型成血管和神经营养效应；明确了黄芪组分对大鼠骨髓源性EPCs的影响及与SCs、HUVEC共培养能够有效促进细胞增殖；证实了黄芪组分体外干预大鼠骨髓源性EPCs局部移植能够有效改善DPN大鼠模型成血管和神经营养效应；为黄芪在糖尿病神经病变的实际应用提供了理论依据。.研究成果已准备在国内核心学术期刊发表（部分论文待见刊）； 培养硕士研究生4名。