中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (16): 2895-2900.doi: 10.3969/j.issn.1673-8225.2011.16.011

• 纳米生物材料 nanobiomaterials • 上一篇    下一篇

移植自聚合肽纳米纤维材料与RhoA-siRNA复合物修复脊髓损伤

江沿文1，张玮玮1，占小多1，康晓宁1，戴  翔2，张文影3，刘忠英1，董为人1，郭家松1   

  1. 1南方医科大学基础医学院组织学与胚胎学教研室，广东省广州市，510515
    2广州达安临床检验中心，广东省广州市，510000
    3广州市奈瑞儿塑身美颜连锁股份有限公司培训学院，广东省广州市 511434
  • 收稿日期:2011-01-14 修回日期:2011-02-28 出版日期:2011-04-16 发布日期:2013-11-11
  • 通讯作者: 郭家松，博士，教授，博士生导师。南方医科大学基础医学院组织学与胚胎学教研室，广东省广州市510515 jiasongguo@yahoo.com.cn
  • 作者简介:江沿文★，1982年生，江西省余干县人，汉族，南方医科大学在读硕士，主要从事中枢神经损伤与修复方面的研究。 jywen1209@163.com
  • 基金资助:

    国家自然科学基金（30973095）资助项目，课题名称：自聚合纳米材料介导siRNA干扰RhoA表达促进脊髓损伤修复的实验研究。

Self-assembling peptide nanofiber scaffold combined with RhoA-siRNA for repair of spinal cord injury

Jiang Yan-wen1, Zhang Wei-wei1, Zhan Xiao-duo1, Kang Xiao-ning1, Dai Xiang2, Zhang Wen-ying3, Liu Zhong-ying1, Dong Wei-ren1, Guo Jia-song1   

  1. 1Department of Histology and Embryology, School of Basic Medical Science, Southern Medical University, Guangzhou  510515, Guangdong Province, China
    2Guangzhou Da'an Clinical Laboratory Center, Guangzhou  510000, Guangdong Province, China
    3Training Institute, Nairuier Body Sculpting and Cosmetic Co., LTD., Guangzhou  511434, Guangdong Province, China
  • Received:2011-01-14 Revised:2011-02-28 Online:2011-04-16 Published:2013-11-11
  • Contact: Guo Jia-song, PhD Professor, Doctoral supervisor, Department of Histology and Embryology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, Guangdong Province, China jiasongguo@yahoo.com.cn
  • About author:Jiang Yan-wen★, Studying for master’s degree, Department of Histology and Embryology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, Guangdong Province, China jywen1209@163.com
  • Supported by:

    National Natural Science Foundation of China, No. 30973095*

PDF

436

被引次数

2

摘要：

背景：脊髓损伤后RhoA表达增高是神经再生困难的主要原因之一，本课题在前期研究证明自聚合肽纳米纤维材料能较好地促进脊髓损伤后结构与功能修复的基础上，构建了自聚合肽纳米纤维材料与RhoA-siRNA复合材料用于修复脊髓损伤。
目的：探讨自聚合肽纳米纤维材料介导siRNA干扰RhoA表达促进脊髓损伤修复。
方法：54只昆明小鼠随机数字表法分为4组。假手术组仅作脊髓暴露，其他3组在切除1 mm脊髓组织制备全横断脊髓损伤模型后，分别于损伤腔内填充生理盐水、自聚合肽纳米纤维支架或含有RhoA特异性siRNA复合物。通过FAM标记检测siRNA转染效率；免疫组化检测RhoA表达；免疫组化和定量分析检测再生神经纤维；行为学检测评定后肢功能恢复。
结果与结论：移植FAM标记的含有RhoA特异性siRNA复合物后，在脊髓内的神经纤维及大脑皮质运动区神经元胞体内均可检测到FAM荧光信号，提示siRNA可从复合材料中释放到组织中并成功导入靶细胞。与SAPNS组和生理盐水组相比，含有RhoA特异性siRNA复合物和自聚合肽纳米纤维支架组可显著降低RhoA在神经元的表达，提高脊髓损伤区内NF阳性神经纤维的密度，促进后肢功能的恢复。结果提示通过自聚合肽纳米纤维支架的介导，含有RhoA特异性siRNA复合物能有效干扰RhoA表达，从而促进脊髓损伤的修复。

关键词: 自聚合肽纳米纤维支架, RhoA, RNA干扰, siRNA, 脊髓损伤, 修复

Abstract:

BACKGROUND: Up-expression of RhoA after spinal cord injury is one of main reasons of neural regeneration failure. Based on our previous report that self-assembling peptide nanofiber scaffold (SAPNS) could effectively promote the repair of structure and function after spinal cord injury, the complex of SAPNS and RhoA-siRNA was constructed to repair the injured spinal cord.
OBJECTIVE: To explore the promotion of SAPNS-mediated siRNA interference RhoA expression on repair of spinal cord injury.
METHODS: A total of 54 Kunming mice were randomly divided into 4 groups: sham group, saline group, SAPNS group and siRNA+SAPNS group. After transection spinal cord injury model was prepared by removed 1mm spinal cord tissue, saline, SAPNS or the complex of siRNA+SAPNS was filled into the lesion cavities in saline group, SAPNS group and siRNA+SAPNS group. Spinal cord was exposed in sham group.Then, the siRNA transfection efficiency was detected by FAM fluorescent signal; the RhoA expression was detected by immnohistochemistry; the axonal regeneration was examined by neurofilament (NF) immnohistochemistry and behavioral test.
RESULTS AND CONCLUSION: After transplantation of FAM-siRNA+SAPNS, the FAM fluorescent signal could be detected in the nerve fiber in the spinal cord and the neuronal bodies in motor cortex, which indicated the siRNA could be released from the complex and successfully entered into the target cells. Compared with the SAPNS group and saline group, RhoA-siRNA+SAPNS transplantation could significantly reduce the RhoA expression in the neurons, increase the densities of NF positive nerve fiber in the spinal cord injury area, and improve the function of the hindlimbs of the spinal cord injured mice. The results indicated that mediated by SAPNS, RhoA-specific siRNA can effectively interfere the expression of RhoA, in order to promote the repair of spinal cord injury.

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