bFGF/PDGF复合水凝胶经PLCγ-AKT信号协同促脊髓损伤修复的研究

Blood spinal cord barrier repair, neuronal protection and axonal regeneration are of great significance to spinal cord injury repair. It is reported that bFGF/PDGF can coordinately promote angiogenesis, Our team previously found that bFGF repairs SCI by promoting BSCB repair and anti-neuronal apoptosis,and that chitosan thermosensitive hydrogel could long-term sustained-release growth factor, We will prepare the bFGF/PDGF complex chitosan thermosensitive hydrogel. The physical and chemical properties of the gel were examined by scanning electron microscopy (SEM) and Elisa method. The effects of in vitro on the growth and function of endothelial and nerve cells, and in vivo on BSCB repair, neuroprotection and axon regeneration are examined using CCK-8 experiment, cell scratch test, skeletal staining, western blot, growth factor in situ tracing, motor function test, BSCB permeability evaluation and nerve repair evaluation to reveal the functions of hydrogel on promoting BSCB and neuroprotection in SCI, clarifying whether bFGF / PDGF synergistic mechanism is related to PLCγ/AKT signaling pathway, and further study the relationship between PLCγ/AKT synergistic signal and autophagy and endoplasmic reticulum stress related proteins. This study provides a new idea for SCI prevention.

血脊髓屏障修复、神经元保护及轴突再生对脊髓损伤修复具有重要的意义。研究报道bFGF/PDGF联用具有协同促血管新生的功能，同时课题组前期发现bFGF通过促血脊髓屏障修复和抗神经元凋亡来修复脊髓损伤以及壳聚糖温敏水凝胶可长期缓释生长因子，我们拟制备载bFGF/PDGF复合壳聚糖温敏水凝胶。利用扫描电镜、Elisa法等检验该凝胶的理化性质，利用CCK-8实验、细胞划痕实验、western blot、生长因子原位示踪、运动功能检查、BSCB渗透性评价、神经修复评价等检验其体外对内皮细胞、神经细胞生长和功能及体内对血脊髓屏障修复、神经保护和轴突再生的影响，并揭示该水凝胶在SCI中协同促BSCB及神经保护的作用，阐明bFGF/PDGF协同机制是否与PLCγ/AKT信号相关，进一步研究PLCγ/AKT协同信号与自噬及内质网应激相关蛋白的关系。该研究对脊髓损伤治疗提供新的思路。

脊髓损伤（SCI）是一种严重的创伤性中枢神经系统疾病，导致神经功能障碍，严重者甚至可致永久性残疾。脊髓损伤后的神经血管再生和运动功能恢复极为困难，也是当前神经科学领域的研究热点。本项目基于自主合成的壳聚糖温敏水凝胶，通过包裹bFGF/PDGF，探索生长因子协同促SCI修复的作用。我们研究显示载bFGF/PDGF水凝胶能够促进脊髓损伤后运动功能和组织损伤的恢复，改善BSCB完整性，抑制细胞凋亡，并发挥内皮细胞保护作用。bFGF/PDGF可能通过协同调控PLCγ/Nrf2/HIF-1α通路，保护内皮细胞，从而发挥BSCB和微循环保护作用，最终促进脊髓损伤后运动神经功能恢复。我们的研究认为，bFGF/PDGF组合对于干预复杂的脊髓损伤后病理过程有巨大的治疗潜力。我们的研究成果为SCI治疗提供了新的治疗策略，在Laboratory Investigation、Oxidative Medicine and Cellular Longevity、Neural Regeneration Research等国际期刊发表SCI15篇，其中第一作者或通讯作者发表论文6篇，授权专利5项。