阿魏酸钠-肝素-泊洛沙姆水凝胶对脊髓损伤大鼠神经修复和再生的影响

Effect of ferulate-heparin-poloxamer thermosensitive hydrogels on nerve regeneration in rats with spinal cord injury

目的 研究阿魏酸钠(sodium ferulate, SF)经肝素-泊洛沙姆(heparin-poloxamer hydrogels, HP)包载后,增强SF对损伤脊髓的修复作用。方法 动物分为假手术组、模型组、空白HP水凝胶组、SF溶液组、SF-HP水凝胶组。术后第1、3、7、14 天行运动能力测试,在d 14,处死大鼠取脊髓行病理染色、检测GFAP、GAP43、Nestin的表达,通过BDA顺行神经示踪法观察神经传导功能。结果 SF-HP水凝胶胶凝温度为37℃,扫描电镜下为三维网状结构;相比模型组,SF-HP组运动功能评分明显升高( P <0.01);HE染色:损伤后出现出血、水肿,SF-HP给药14 d后,能改善此现象;蛋白检测:相比SF组,SF-HP给药后GAP43、Nestin表达增加( P <0.05),GFAP表达下降( P <0.05)。此外,BDA示踪显示,SF水凝胶给药组阳性神经元数目明显增加( P <0.05),神经传导功能修复明显。结论 通过水凝胶载体的缓释作用,增强了SF在受损脊髓处神经功能重建及神经再生的作用效果。

Aim To investigate the effect of ferulate-heparin-poloxamer(SF-HP) thermosensitive hydrogels on nerve regeneration in ratswith spinal cord injury, and provide experimental basis for clinical application. Methods SD rats were randomly divided into five groups: sham group, SCI group, SCI+HP group, SCI+SF group and SCI+SF-HP group. Rats were performed moderate contusion injuries using a vascular clip for 2 min to establish SCI animal model, then rats were given BBB score and inclined plate scoring function test after SCI. BDA tracing was employed to observe the recovery of nerve conduction function. Moreover, immunohistochemical staining and Western blotwere used to measure GAP43, Nestin and GFAP levels. Results BBB score and inclined plane test score significantly increased in SF-HP treated group as compared with the model group( P <0.01). Staining data showed that the structure of spinal cord was void, and this phenomenon was reversed after SF-HP administration. Data showed that the level of GAP43 and Nestin increased after SF treatment( P <0.05), the expression of GFAP decreased( P <0.05), and the treatment effect of SF-HP hydrogels were better than those of SF alone( P <0.05). BDA tracing data showed that the number of positive neurons markedly increased and the repair of nerve conduction function was significantly improved in SF-HP hydrogel group compared with SF group. Conclusion HP hydrogels enhance the effect of SF on the nerve regeneration in damaged spinal cord in rats.