Combined Treatment of Neurotrophin-3 Gene and Neural Stem Cells is Propitious to Functional Recovery after Spinal Cord Injury
Combined Treatment of Neurotrophin-3 Gene and Neural Stem Cells is Propitious to Functional Recovery after Spinal Cord Injury
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DOI：
10.3727/000000007783464902
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发表时间：
2007-05
影响因子：
3.3
通讯作者：
Lu-min Zhang;Shuting Gu;Cui-ping Zhao;T. Wen
中科院分区：
文献类型：
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作者：
Lu-min Zhang;Shuting Gu;Cui-ping Zhao;T. Wen
我们介绍了神经营养因子-3和神经干细胞联合治疗对脊髓损伤(SCI)后功能恢复的影响。从神经干细胞系C17.2中提取总RNA，逆转录成cDNA。用聚合酶链式反应扩增神经营养素-3(NT-3)基因，将其亚克隆到载体pNT-3中，构建表达载体pNT-3。获得了含有PNT-3的阳性克隆，命名为SHN2，并用于移植。30只成年小鼠在T8椎体水平接受机械性损伤。X-Gal染色、RT-PCR和旷场运动分别观察细胞存活、NT-3基因表达和功能恢复情况。结果表明，克隆了长777个核苷酸的NT-3基因，并将其导入神经干细胞系C17.2，获得了2倍以上的表达。细胞密度的定量分析显示，与对照细胞相比，SHN2细胞显著增加(p<；0.01)。移植后30天，SHN_2在病损部位附近显著增加。此外，功能恢复通过Basso-Beattie-Bresnahan(BBB)运动评定量表(p<；0.01)检测显示有积极作用。结论：神经干细胞与NT-3基因联合治疗可促进神经功能恢复。它为治疗脊髓损伤提供了一种有效的方法。
We present an insight of the effects of combination therapy with neurotrophin-3 and neural stem cell on functional recovery after spinal cord injury (SCI). Total RNA was extracted from neural stem cell line C17.2 and reversed transcribed into cDNA. Neurotrophin-3 (NT-3) gene was amplified by PCR and subcloned into plasmid to construct an expression vector pNT-3. A positive clone containing pNT-3, named SHN2, was obtained and used for transplantation. Thirty adult mice received mechanical injury at the T8 vertebra level. Cell survival, NT-3 gene expression, and functional recovery were observed through X-Gal staining, RT-PCR, and open field locomotion, respectively. The results show that NT-3 gene comprising 777 bp nucleo-tides was cloned and a more than twofold expression was detected when transfected into neural stem cell line C17.2. Quantitative analysis of cellular density revealed a significant increase in SHN2 compared to the control cells (p < 0.01). Thirty days after transplantation, SHN2 showed significant increase near the lesion site. Furthermore, the functional recovery indicated an active effect by detecting Basso-Beattie-Bresnahan (BBB) locomotor rating scale (p < 0.01). In conclusion, combined treatment of neural stem cells and NT-3 gene can facilitate functional recovery. It offers an effective approach to treat SCI.