硫酸软骨素蛋白聚糖（CSPGs）结合肽联合激动型L1抗体治疗脊髓损伤实验研究

项目名称： 硫酸软骨素蛋白聚糖（CSPGs）结合肽联合激动型L1抗体治疗脊髓损伤实验研究

项目编号： No.81471279

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 赵炜疆

作者单位： 汕头大学

项目金额： 70万元

中文摘要： 脊髓损伤后神经元再生是巨大的医学难题。损伤部位胶质瘢痕中有益分子相对匮乏，而修复抑制分子如硫酸软骨素蛋白聚糖（CSPGs）严重阻碍损伤后功能恢复。我们已有研究结果表明小鼠脊髓损伤后给予激动型细胞黏附分子L1抗体可有效促进神经元轴突延伸进入胶质瘢痕，但不能有效穿越损伤断端。封闭CSPGs功能位点，抑制CSPGs作用，进一步激活一系列与损伤修复有关的信号级联反应，可能成为促进脊髓损伤后结构重建和功能恢复的重要环节。本课题拟首先通过噬菌体展示技术筛选具有生物活性的CSPGs特异性结合肽，并综合使用细胞生物学、分子生物学、形态学以及行为学等实验方法，观察单独使用结合肽以及结合肽与激动型细胞黏附分子L1抗体联用对脊髓损伤小鼠神经再生、胶质瘢痕以及运动功能改善的影响，确立以促进轴突再生和中和修复抑制因子为主导的脊髓损伤联合治疗方案，为临床应用提供有益参考。

中文关键词： 脊髓损伤；神经再生；硫酸软骨素蛋白聚糖；结合肽；噬菌体展示

英文摘要： Spinal cord injury is a detrimentally clinical setting due to the difficult neuronal regeneration as a large medical challenge. The lackness of beneficial molecules and the accumulation of regeneration inhibition molecules chondroitin sulfate proteoglycans (CSPGs) severely abrogate the functional recovery post injury. The data available from our experiments demonstrated that administration of stimulatory domain antibody targeting cell adhesion molecule L1 can effectively promote the elongation of neuronal axons into the glial scar, while unsuccessful crossing to the site distal to the injury was observed. Thus, inhibiton of CSPGs function by neutralizing the functional epitope of CSPGs may stimulate a series of signaling cascades related to neuronal regeneration, thus representing a key step of structural reconstruction and functional recovery after spinal cord injury. In the present project, we purposed to apply phage display (PD) to screen the specific binding peptide for CSPGs with a strong biological activity. Then, PCR, Western blot (WB)， immunohistochemistry (IHC)， immunofluorescence (IF), and behavioral methods were employed to study the single use of the binding peptide, or the combinatorial application of binding peptide and stimulatory antibody targeting the cell adhesion molecule L1, on post-injury neuronal regeneration, glial scar formation, as well as on motion improvement. Through these experimentl designs, we hope to establish a therapeutic regimen composed of stimulating neuronal regeneration and neutralizing regeneration inhibition molecules. This project may provide a valuable clue for clinical management of spinal cord injury.

英文关键词： spinal cord injury;neuronal regeneration;chondroitin sulfate proteoglycan (CSPGs);binding peptide;phage display