Effect of exogenous spastin combined with polyethylene glycol on sciatic nerve injury
Effect of exogenous spastin combined with polyethylene glycol on sciatic nerve injury
复制标题
外源性spastin联合聚乙二醇对坐骨神经损伤的影响
DOI：
10.4103/1673-5374.251336
复制
发表时间：
2019-07-01
影响因子：
6.1
通讯作者：
Lin, Hao-Dong
中科院分区：
文献类型：
--
作者：
Lin, Yao-Fa;Xie, Zheng;Lin, Hao-Dong
聚乙二醇能通过轴突融合在细胞水平将损伤神经的远端和近端连接起来，避免损伤远端神经的沃勒氏变性，促进周围神经再生。然而，由于细胞骨架没有得到及时修复，这种方法只能防止10%的轴突发生沃勒氏变性。重建细胞骨架干和微管网络被认为是提高轴突融合效率的关键。作为一种切断微管的蛋白质，spastin已被用于促进细胞骨架的重建。因此，我们推测SPASTIN联合聚乙二醇能更有效地促进周围神经再生。雄性SD大鼠120只，随机分为假手术组、缝合组、聚乙二醇组、聚乙二醇组和聚乙二醇组。缝合组大鼠坐骨神经切断后仅行传统的端端缝合神经吻合。聚乙二醇组和聚乙二醇组分别在远端缝合神经外膜下立即注射50μ的聚乙二醇组L或25μ的聚乙二醇组L+25μ的巴斯汀组。用来评价术后1周早期神经再生的感觉纤维再生距离，缝合组最短，聚乙二醇组次之，聚乙二醇组最大。大鼠运动功能恢复的行为学评估显示，术后8周，聚乙二醇组和聚乙二醇组+痉挛素组肢体功能均恢复正常。术后1，2，4，8周，假手术组坐骨神经功能指数和腓肠肌湿重百分比最高，其次为聚乙二醇组和聚乙二醇组，缝合组最低。用Masson染色评价肌肉组织的形态。术后1、2、4、8周，骨缝线组、聚乙二醇组和聚乙二醇组均可检测到骨骼肌的形态变化。其中，缝合组肌萎缩最为严重，其次为聚乙二醇组和聚乙二醇组+痉挛素组。术后1周、2周、4周、8周，远侧坐骨神经组织超微结构显示缝线组、聚乙二醇组和聚乙二醇组在术后1周、2周、4周、8周出现早期破坏、继而崩解、逐渐修复的形态。随着时间的推移，轴突超微结构逐渐恢复。事实上，术后8周时，聚乙二醇组和假手术组相似。我们的研究结果表明，聚乙二醇和痉挛蛋白联合应用可以促进周围神经再生。且联合应用效果优于单纯聚乙二醇组，且均优于传统神经缝合术。本研究由第二军医大学动物伦理委员会中国(批准号：CZ20170216)2017年3月16日。
Polyethylene glycol can connect the distal and proximal ends of an injured nerve at the cellular level through axonal fusion to avoid Wallerian degeneration of the injured distal nerve and promote peripheral nerve regeneration. However, this method can only prevent Wallerian degeneration in 10% of axons because the cytoskeleton is not repaired in a timely fashion. Reconstruction of the cytoskeletal trunk and microtubule network has been suggested to be the key for improving the efficiency of axonal fusion. As a microtubule-severing protein, spastin has been used to enhance cytoskeletal reconstruction. Therefore, we hypothesized that spastin combined with polyethylene glycol can more effectively promote peripheral nerve regeneration. A total of 120 male Sprague-Dawley rats were randomly divided into sham, suture, polyethylene glycol, and polyethylene glycol + spastin groups. In suture group rats, only traditional nerve anastomosis of the end-to-end suture was performed after transection of the sciatic nerve. In polyethylene glycol and polyethylene glycol + spastin groups, 50 μL of polyethylene glycol or 25 μL of polyethylene glycol + 25 μL of spastin, respectively, were injected immediately under the epineurium of the distal suture. Sensory fiber regeneration distance, which was used to assess early nerve regeneration at 1 week after surgery, was shortest in the suture group, followed by polyethylene glycol group and greatest in the polyethylene glycol + spastin group. Behavioral assessment of motor function recovery in rats showed that limb function was restored in polyethylene glycol and polyethylene glycol + spastin groups at 8 weeks after surgery. At 1, 2, 4 and 8 weeks after surgery, sciatic functional index values and percentages of gastrocnemius muscle wet weight were highest in the sham group, followed by polyethylene glycol + spastin and polyethylene glycol groups, and lowest in the suture group. Masson staining was utilized to assess the morphology of muscle tissue. Morphological changes in skeletal muscle were detectable in suture, polyethylene glycol, and polyethylene glycol + spastin groups at 1, 2, 4, and 8 weeks after surgery. Among them, muscular atrophy of the suture group was most serious, followed by polyethylene glycol and polyethylene glycol + spastin groups. Ultrastructure of distal sciatic nerve tissue, as detected by transmission electron microscopy, showed a pattern of initial destruction, subsequent disintegration, and gradual repair in suture, polyethylene glycol, and polyethylene glycol + spastin groups at 1, 2, 4, and 8 weeks after surgery. As time proceeded, axonal ultrastructure gradually recovered. Indeed, the polyethylene glycol + spastin group was similar to the sham group at 8 weeks after surgery. Our findings indicate that the combination of polyethylene glycol and spastin can promote peripheral nerve regeneration. Moreover, the effect of this combination was better than that of polyethylene glycol alone, and both were superior to the traditional neurorrhaphy. This study was approved by the Animal Ethics Committee of the Second Military Medical University, China (approval No. CZ20170216) on March 16, 2017.