中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (36): 6751-6754.doi: 10.3969/j.issn.1673-8225.2010.36.022

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

神经营养因子3基因修饰神经干细胞移植脊髓损伤的相关蛋白表达

邓兴力1,梁袁昕2,杨智勇1,宋晓斌1,李  玉1,杨述华2   

  1. 1昆明医学院第一附属医院神经外科,云南省昆明市  650032;2华中科技大学同济医学院协和医院骨科, 湖北省武汉市  430022
  • 出版日期:2010-09-03 发布日期:2010-09-03
  • 作者简介:邓兴力☆,男,1979年生,云南省昆明市人,汉族,2008年昆明医学院毕业,博士,讲师,主要从事神经干细胞与神经再生研究。 duetch13071@ yahoo.com.cn

Spinal cord injury-associated protein expression following neurotrophin-3 gene modified neural stem cells transplantation  

Deng Xing-li1, Liang Yuan-xin2, Yang Zhi-yong1, Song Xiao-bin1, Li Yu1, Yang Shu-hua2   

  1. 1 Department of Neurosurgery, First Affiliated Hospital, Kunming Medical College, Kunming  650032, Yunnan Province, China;2 Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
  • Online:2010-09-03 Published:2010-09-03
  • About author:Deng Xing-li☆, Doctor, Lecturer, Department of Neurosurgery, First Affiliated Hospital, Kunming Medical College, Kunming 650032, Yunnan Province, China duetch13071@ yahoo.com.cn

摘要:

背景:研究表明神经干细胞和神经营养因子3基因修饰的神经细胞联合移植能够在移植后存活并有效促进脊髓横断后脊髓的功能恢复,但神经营养因子3基因修饰的神经干细胞能否在脊髓受损部位发挥功能并促进脊髓损伤大鼠的功能恢复?
目的:观察神经营养因子3基因修饰胚胎脊髓神经干细胞移植后脊髓损伤大鼠的功能恢复情况及损伤局部的基因表达。
方法:将30只SD大鼠在T9水平进行脊髓半切后,随机分为3组,分别在受损脊髓内植入细胞培养液、神经干细胞及神经营养因子3基因修饰神经干细胞。另取10只仅行椎板切除设置为空白对照。移植后通过行为学测试评价脊髓功能的恢复,RT-PCR和Western blot检测脊髓损伤部位神经营养因子3和髓鞘碱性蛋白的表达。
结果与结论:移植神经营养因子3基因修饰神经干细胞组行为学测试结果最好,移植细胞培养液组行为学测试最差。与移植细胞培养液组相比,移植神经干细胞及神经营养因子3基因修饰神经干细胞组大鼠脊髓组织中神经营养因子3基因和髓鞘碱性蛋白基因的mRNA水平明显上调,在蛋白水平也有类似的结果,且神经营养因子3基因修饰神经干细胞组效果更明显。提示移植神经营养因子3基因修饰神经干细胞能促进脊髓受损部位出现更多向少突胶质细胞分化的细胞,并能更强的表达神经营养因子3。

关键词: 神经干细胞, 神经营养因子3, 移植, 脊髓损伤, 干细胞移植

Abstract:

BACKGROUND: Studies have confirmed that neural stem cells (NSCs) and neurotrophic factor-3 (NT-3) gene modified neural cell combination transplantation can survive following transplantation and effectively promote functional recovery following spinal cord transsection. However, whether NT-3 gene-modified NSCs exert effects in damaged spinal cord and contribute to functional recovery of the spinal cord injury (SCI) rats remains poorly understood. OBJECTIVE: To investigate the behavioral recovery of SCI rats that received transplantation of NT-3 gene-modified fetal spinal cord-derived NSCs, and the gene expression in injury location.
METHODS: Spinal cords of 30 Sprague-Dawley rats were hemisected at T9 level. The rats were randomly assigned to three groups. Cell culture medium, NSCs and NT-3 gene-modified NSCs were transplanted into the lesion site of rats of each group respectively. An additional 10 rats served as blank control group, which only received laminectomy. Following transplantation, behavior test was utilized to evaluate spinal cord functional recovery. Reverse transcription-polymerase chain reaction and Western blot assay were used to detect NT-3 and myelin basic protein expression in lesion site.
RESULTS AND CONCLUSION: Rats in NT-3 gene -NSCs group got best results in behavioral test. Behavioral test outcomes were worst in the cell culture medium group. Compared with cell culture medium group, NT-3gene and myelin basic protein gene mRNA expression significantly upregulated in spinal cord tissue of rats from the NSCs and NT-3 gene-modified NSCs groups. Protein levels obtained similar results. Moreover, the outcomes were obvious in the NT-3 gene -modified NSCs group. These indicate that transplantation of NT-3 gene -modified NSCs can promote the differentiation of cells into oligodendrocytes in lesion site, and strongly express NT-3.

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