基于调节神经干细胞增殖分化的振荡电场刺激修复脊髓损伤及机制研究

Neural stem cell transplantation increases the hope for treatment of spinal cord injury, but it still exists a problem that the proliferation and differentiation of neural stem cell is interfered by the microenvironment in the injured spinal cord. A single neural stem cell transplantation can not meet the need for treatment of spinal cord injury. It has shown that the proliferation and differentiation of neural stem cell is promoted by the Wnt signaling pathway in previous studies. The electrical field stimulation is an effective physical method for the treatment of spinal cord injury proved by clinical trials, and it can activate the Wnt signaling pathway in the microenvironment of spinal cord. On this basis, the purpose of this project is to repair the injured spinal cord in rats by combining oscillating field stimulation and neural stem cell transplantation. Observing the proliferation and differentiation of exogenous neural stem cell by immunofluorescence labeling technology ; detecting the expression and distribution of Wnt signaling pathway protein in the injured spinal cord by qRT-PCR and Western blot ; observing the regeneration of axons and myelin by tissue staining and electron microscope technology; evaluating the functional recovery via BBB score and neurophysiological test. Exploring the mechanism of inducing the proliferation and differentiation of neural stem cells and promoting neurons regeneration by oscillating field stimulation; confirming the hypothesis that the oscillating electric field stimulation can promote the proliferation and differentiation of neural stem cell; Providing a theoretical basis and a new direction for the treatment of spinal cord injury by means of oscillating electric field and neural stem cell transplantation.

神经干细胞移植是治疗脊髓损伤的重要手段，但神经干细胞移植存在增殖和分化易受到脊髓微环境干扰的问题。Wnt信号通路对调控干细胞分化及促进神经再生具有重要作用。振荡电场刺激是经临床试验证明有效的修复脊髓损伤的物理方法，可调控Wnt信号通路。本研究采用大鼠脊髓损伤模型，联合应用振荡电场刺激和神经干细胞移植修复损伤脊髓，利用免疫荧光标记、qRT-PCR、Western blot、免疫荧光染色、基因芯片和动物行为学等技术，观察振荡电场刺激对移植的神经干细胞增殖分化和神经再生的调节作用；分析Wnt蛋白的表达及其下游信号通路的激活，探讨在振荡电场刺激下，参与Wnt信号通路调节神经干细胞增殖分化的机制；并进一步研究抑制或增强脊髓内Wnt信号通路对神经再生的影响。以证实振荡电场刺激调节移植的神经干细胞增殖分化和促进神经再生的假说，为联合应用振荡电场刺激和神经干细胞移植修复脊髓损伤提供新的理论依据和技术方向。

脊髓损伤是一种常见的严重创伤，神经干细胞移植是治疗脊髓损伤的重要手段，但神经干细胞移植存在增殖和分化易受到脊髓微环境干扰的问题。Wnt 信号通路对调控干细胞分化及促进神经再生具有重要作用。振荡电场刺激是经临床试验证明有效的修复脊髓损伤的物理方法，可调控 Wnt 信号通路。本课题采用大鼠脊髓损伤模型，应用振荡电场刺激修复损伤脊髓，利用Western blot、免疫荧光染色、动物行为学等技术，观察振荡电场刺激对内源性神经干细胞增殖分化和神经再生的调节作用；分析 Wnt 蛋白的表达及其下游信号通路的激活，探讨在振荡电场刺激下，参与 Wnt 信号通路调节神经干细胞增殖分化的机制。结果表明振荡电场可以促进 Wnt-3a、β-catenin 的高表达来促进内源性神经干细胞的增殖和分化，从而达到治疗脊髓损伤的目的。通过使用表面电刺激治疗临床脊髓损伤患者，发现表面电刺激治疗可显著改善脊髓损伤后逼尿肌痉挛型膀胱患者尿流动力学指标和膀胱功能，且对颈、胸髓损伤患者的疗效优于腰髓损伤患者。项目组同时分析评估了小鼠脊髓挫伤后运动皮层的经颅视觉运动标测，结果表明，经颅视觉运动标测是评价不同程度脑挫裂伤后结构和功能损伤及可塑性的一种有效、灵敏的方法。综合上述结果，本研究为探讨修复脊髓损伤的新策略奠定了坚实的工作基础。项目资助发表论文4篇，培养硕士研究生4名，其中2名已经取得硕士学位，2名在读。