SHH缓释的脊髓去细胞支架促进大鼠脊髓损伤修复  

Release of SHH by acellular spinal cord scaffold promotes function recovery after spinal cord injury in rats

作　　者：陈平波[1,2] 徐晓峰 周焕高[1,2] 毕士奇 史文涛 杨开元 张志坚 陈谦[2] 杨文静[2] Chert Pingbo;Xu Xiaofeng;Zhou Huangao;Bi shiqi;Shi Wentao;Yang Kaiyuan;Zhang Zhijian;Chert Qian;Yang Wenjing(Department of orthopedics, Affiliated Hospital;School of Medicine, Jiangsu University, Zhenjiang 212001, China)

机构地区：[1]江苏大学附属医院骨科,镇江212001 [2]江苏大学医学院,镇江212001

出　　处：《神经解剖学杂志》2018年第2期203-209,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81501077;81571830;51403086);江苏省普通高校研究生科研创新计划项目(SJZZ16-0202;SJLX16-0449)

摘　　要：目的:研究可缓释音猬因子(sonic hedgehog,SHH)的脊髓去细胞支架(acellular spinal cord scaffolds,ASCSs)对大鼠脊髓损伤修复的效果。方法:(1)用物理和化学联合法制备ASCSs,并检测其效果。(2)使用京尼平(genipin,GP)作为交联剂,体外制备缓释SHH的ASCSs(ASCSs-GP-SHH),探究其缓释效果。(3)建立大鼠脊髓损伤(spinal cord injury,SCI)模型,随机分为四组(每组20只):单纯SCI组,ASCSs组,ASCSs-GP组(GP交联的ASCSs),ASCSs-GP-SHH组。术后每周记录BBB评分,评估功能恢复。术后12周取出损伤处脊髓组织,行免疫组化和Western Blot检测损伤处相关蛋白Nestin(巢蛋白),GAP43(生长相关蛋白),MBP(髓磷脂碱性蛋白),NF200(神经丝蛋白),GFAP(胶质纤维酸性蛋白)的表达。结果:(1)物理及化学联法制备ASCSs效果良好;(2)ASCSs-GP-SHH具有良好的缓释SHH的性能;(3)ASCS-GP-SHH组BBB评分从术后到12周较其他组高(P<0.05);(4)免疫组化及HE染色结果显示:ASCS-GP-SHH组脊髓横断处有神经元再生。SCI组神经元凋亡明显,纤维瘢痕及空洞大,ASCS-GP组较ASCS组稍好,ASCS组较SCI组稍好;(5)Western Blot结果显示:ASCS-GP-SHH组Nestin,GAP43,MBP,NF200表达高于SCI、ASCS-GP、ASCS组(P<0.05),而GFAP低于SCI、ASCS-GP、ASCS组(P<0.05)。结论:物理和化学联合制备可以有效的制备ASCSs,ASCSs-GP-SHH缓释效果良好,对大鼠脊髓损伤的修复有明显的促进作用。Objective： To study the effect of acellular spinal cord scaffolds（ ASCSs） that sustain release of SHH in the repair of spinal cord injury in rats. Methods：（ 1） Preparation of ASCSs by using physical and chemical methods,and evaluate the efficacy of decellularization;（ 2） Preparation for ASCSs that can sustain release of SHH by using genipin as a crosslinking agent,and determination of SHH release from ASCSs-GP-SHH.（ 3） The spinal cord injury model was established,the rat were randomly divided into four groups（ each group of 20） ： SCI group; ASCSs group; ASCSsGP group; ASCSs-GP-SHH group. BBB scores were recorded weekly to assess functional recovery. After 12 weeks,the lesioned area tissue was oollected,immunohistochemistry and Western Blot were used to detect the espression of Nestin,GAP43,MBP,NF200 and GFAP. Results：（ 1） The preparation of ASCSs by physical and chemical method was effi-cacy.（ 2） ASCSs-GP-SHH has a good performance of sustained-release of SHH.（ 3） After 12 weeks,The BBB score of ASCS-GP-SHH group was higher than that of other groups（ P〈0. 05）.（ 4） Immunohistochemistry and HE staining showed that there was neuronal regeneration in the spinal cord of ASCS-GP-SHH group. SCI group showed prominent apoptosis and marked tiber scar and vacuola. In these physiological apects,the sererity decreased in the following order among groups; ASCSGP,ASCS and SCI.（ 5） Western Blot showed that the expression of Nestin,GAP43,MBP and NF200 in ASCS-GP-SHH group was higher than that in SCI,ASCS-GP and ASCS（ P〈0. 05）,while GFAP was lower than that of SCI,ASCS-GP and ASCS（ P〈0. 05）. Conclusion： The ASCSs prepared by combination of physical and chemical are effective,and the sustained release of ASCSs-GP-SHH can promote the repair of spinal cord injury in rats.

关 键 词：音猬因子 脊髓去细胞支架 京尼平 缓释 脊髓损伤 

分 类 号：R651.2[医药卫生—外科学]