Effect of <i>NEP1</i>-<i>40</i> Gene Modified Neural Stem Cell Transplantation on the Behavior Recovery of Rats after Spinal Cord Injury_West China Medical Journal

Authors：

 WANGLei ,  SONGYue-ming , LIULi-min , LITao , GONGQuan , YANGXi

Department of Orthopedics, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P. R. China;

Corresponding author：

SONGYue-ming, Email: sym_cd@yahoo.com.cn

Keywords：

NEP1-40 gene; Neural stem cell transplantation; Spinal cord injury; Basso-Beattle-Bresnahan test; Grid test; Rats

DOI：

10.7507/1002-0179.20140609

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the behavioral recovery of spinal cord injury (SCI) rats that received transplantation of NEP1-40 gene-modified neural stem cells. Methods Neural stem cells (NSCs) were derived from the cortex tissue of rat embryo at the age of 18 days and identified by Nestin immunofluorescence. The lentiviruses were transduced to NSCs to construct NEP1-40 gene modified NSCs. Spinal cords of 30 Sprague-Dawley rats were hemisected at the nineth thoracic vertebrae level. The rats were randomly assigned to three groups. Cell culture medium, NSCs and NEP1-40 gene-modified NSCs were transplanted into the lesion site of rats of SCI group, NSCs group and NEP1-40-NSCs group respectively 7 days after injury. Additional 10 rats served as blank control group (sham group), which only received laminectomy. Following transplantation, behavior tests including Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale and grid test were utilized to evaluate spinal cord functional recovery. Results Behavior tests 8 weeks after cells transplantation showed that the rats in SCI group got worst results, the BBB scores improved and the grid drop times reduced significantly in NSCs transplantation group (P<0.01) and behavioral test outcomes were best in the NEP1-40 gene-modified NSCs group (P<0.01). Conclusion NEP1-40 gene modification can significantly improve the behavioral recovery of SCI rats that received transplantation of pure neural stem cells. It can provide a new idea and reliable experimental base for the study of NSCs transplantation for spinal cord injury.

Citation： WANGLei, SONGYue-ming, LIULi-min, LITao, GONGQuan, YANGXi. Effect of NEP1-40 Gene Modified Neural Stem Cell Transplantation on the Behavior Recovery of Rats after Spinal Cord Injury. West China Medical Journal, 2014, 29(11): 2006-2011. doi: 10.7507/1002-0179.20140609 Copy

1.	Kabatas S, Teng YD. Potential roles of the neural stem cell in the restoration of the injured spinal cord:review of the literature[J]. Turk Neurosurg, 2010, 20(2):103-110.
2.	Steward O, Sharp K, Yee KM, et al. A re-assessment of the effects of a Nogo-66 receptor antagonist on regenerative growth of axons and locomotor recovery after spinal cord injury in mice[J]. Exp Neurol, 2008, 209(2):446-468.
3.	Atalay B, Bavbek M, Ozen O, et al. Nogo-A inhibitory peptide (NEP1-40) increases pan-cadherin expression following mild cortical contusion injury in rats[J]. Turk Neurosurg, 2008, 18(4):356-365.
4.	朱薇薇, 赵红洋, 温天莲, 等. 缺氧缺血性脑损伤新生大鼠脑Nogo受体水平及NEP1-40的神经保护作用[J]. 中华儿科杂志, 2010, 48(2):138-142.
5.	Wang Q, Gou X, Xiong L, et al. Trans-activator of transcription-mediated delivery of NEP1-40 protein into brain has a neuroprotective effect against focal cerebral ischemic injury via inhibition of neuronal apoptosis[J]. Anesthesiology, 2008, 108(6):1071-1080.
6.	袁海峰, 宋跃明, 刘浩, 等. NEP1-40基因慢病毒载体构建及鉴定[J]. 中国修复重建外科杂志, 2012, 26(2):177-181.
7.	Li Y, Zhang WM, Wang TH. Optimal location and time for neural stem cell transplantation into transected rat spinal cord[J]. Cell Mol Neurobiol, 2011, 31(3):407-414.
8.	Mcmahon SS, Albermann S, Rooney GE, et al. Engraftment, migration and differentiation of neural stem cells in the rat spinal cord following contusion injury[J]. Cytotherapy, 2010, 12(3):313-325.
9.	Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats[J]. J Neurotrauma, 1995, 12(1):1-21.
10.	Metz GA, Merkler D, Dietz V, et al. Efficient testing of motor function in spinal cord injured rats[J]. Brain Res, 2000, 883(2):165-177.
11.	Fawcett J. Repair of spinal cord injuries:where are we, where are we going?[J]. Spinal Cord, 2002, 40(12):615-623.
12.	Fournier AE, Grandpre T, Strittmatter SM. Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration[J]. Nature, 2001, 409(6818):341-346.
13.	Chen MS, Huber AB, Van Der Haar ME, et al. Nogo-A is a myelin-associated neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-1[J]. Nature, 2000, 403(6768):434-439.
14.	Williams G, Wood A, Williams EJ, et al. Ganglioside inhibition of neurite outgrowth requires Nogo receptor function:identification of interaction sites and development of novel antagonists[J]. J Biol Chem, 2008, 283(24):16641-16652.
15.	Su Z, Cao L, Zhu Y, et al. Nogo enhances the adhesion of olfactory ensheathing cells and inhibits their migration[J]. J Cell Sci, 2007, 120(Pt 11):1877-1887.
16.	Hou T, Shi Y, Cheng S, et al. Nogo-A expresses on neural stem cell surface[J]. Int J Neurosci, 2010, 120(3):201-205.
17.	Grandpré T, Li S, Strittmatter SM. Nogo-66 receptor antagonist peptide promotes axonal regeneration[J]. Nature, 2002, 417(6888):547-551.
18.	Li S, Strittmatter SM. Delayed systemic Nogo-66 receptor antagonist promotes recovery from spinal cord injury[J]. J Neurosci, 2003, 23(10):4219-4227.
19.	Bart J, Groen HJ, Hendrikse NH, et al. The blood-brain barrier and oncology:new insights into function and modulation[J]. Cancer Treat Rev, 2000, 26(6):449-462.
20.	宫福良, 王坤正, 余鹏博, 等. NEP1-40基因克隆及蛋白的原核表达和纯化[J]. 中国修复重建外科杂志, 2006, 20(1):9-12.
21.	Basso DM, Beattie MS, Bresnahan JC, et al. MASCIS evaluation of open field locomotor scores:effects of experience and teamwork on reliability. Multicenter Animal Spinal Cord Injury Study[J]. J Neurotrauma, 1996, 13(7):343-359.
22.	Z'graggen WJ, Metz GA, Kartje GL, et al. Functional recovery and enhanced corticofugal plasticity after unilateral pyramidal tract lesion and blockade of myelin-associated neurite growth inhibitors in adult rats[J]. J Neurosci, 1998, 18(12):4744-4757.
23.	Metz GA, Schwab ME, Welzl H. The effects of acute and chronic stress on motor and sensory performance in male Lewis rats[J]. Physiol Behav, 2001, 72(1/2):29-35.

1. Kabatas S, Teng YD. Potential roles of the neural stem cell in the restoration of the injured spinal cord:review of the literature[J]. Turk Neurosurg, 2010, 20(2):103-110.
2. Steward O, Sharp K, Yee KM, et al. A re-assessment of the effects of a Nogo-66 receptor antagonist on regenerative growth of axons and locomotor recovery after spinal cord injury in mice[J]. Exp Neurol, 2008, 209(2):446-468.
3. Atalay B, Bavbek M, Ozen O, et al. Nogo-A inhibitory peptide (NEP1-40) increases pan-cadherin expression following mild cortical contusion injury in rats[J]. Turk Neurosurg, 2008, 18(4):356-365.
4. 朱薇薇, 赵红洋, 温天莲, 等. 缺氧缺血性脑损伤新生大鼠脑Nogo受体水平及NEP1-40的神经保护作用[J]. 中华儿科杂志, 2010, 48(2):138-142.
5. Wang Q, Gou X, Xiong L, et al. Trans-activator of transcription-mediated delivery of NEP1-40 protein into brain has a neuroprotective effect against focal cerebral ischemic injury via inhibition of neuronal apoptosis[J]. Anesthesiology, 2008, 108(6):1071-1080.
6. 袁海峰, 宋跃明, 刘浩, 等. NEP1-40基因慢病毒载体构建及鉴定[J]. 中国修复重建外科杂志, 2012, 26(2):177-181.
7. Li Y, Zhang WM, Wang TH. Optimal location and time for neural stem cell transplantation into transected rat spinal cord[J]. Cell Mol Neurobiol, 2011, 31(3):407-414.
8. Mcmahon SS, Albermann S, Rooney GE, et al. Engraftment, migration and differentiation of neural stem cells in the rat spinal cord following contusion injury[J]. Cytotherapy, 2010, 12(3):313-325.
9. Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats[J]. J Neurotrauma, 1995, 12(1):1-21.
10. Metz GA, Merkler D, Dietz V, et al. Efficient testing of motor function in spinal cord injured rats[J]. Brain Res, 2000, 883(2):165-177.
11. Fawcett J. Repair of spinal cord injuries:where are we, where are we going?[J]. Spinal Cord, 2002, 40(12):615-623.
12. Fournier AE, Grandpre T, Strittmatter SM. Identification of a receptor mediating Nogo-66 inhibition of axonal regeneration[J]. Nature, 2001, 409(6818):341-346.
13. Chen MS, Huber AB, Van Der Haar ME, et al. Nogo-A is a myelin-associated neurite outgrowth inhibitor and an antigen for monoclonal antibody IN-1[J]. Nature, 2000, 403(6768):434-439.
14. Williams G, Wood A, Williams EJ, et al. Ganglioside inhibition of neurite outgrowth requires Nogo receptor function:identification of interaction sites and development of novel antagonists[J]. J Biol Chem, 2008, 283(24):16641-16652.
15. Su Z, Cao L, Zhu Y, et al. Nogo enhances the adhesion of olfactory ensheathing cells and inhibits their migration[J]. J Cell Sci, 2007, 120(Pt 11):1877-1887.
16. Hou T, Shi Y, Cheng S, et al. Nogo-A expresses on neural stem cell surface[J]. Int J Neurosci, 2010, 120(3):201-205.
17. Grandpré T, Li S, Strittmatter SM. Nogo-66 receptor antagonist peptide promotes axonal regeneration[J]. Nature, 2002, 417(6888):547-551.
18. Li S, Strittmatter SM. Delayed systemic Nogo-66 receptor antagonist promotes recovery from spinal cord injury[J]. J Neurosci, 2003, 23(10):4219-4227.
19. Bart J, Groen HJ, Hendrikse NH, et al. The blood-brain barrier and oncology:new insights into function and modulation[J]. Cancer Treat Rev, 2000, 26(6):449-462.
20. 宫福良, 王坤正, 余鹏博, 等. NEP1-40基因克隆及蛋白的原核表达和纯化[J]. 中国修复重建外科杂志, 2006, 20(1):9-12.
21. Basso DM, Beattie MS, Bresnahan JC, et al. MASCIS evaluation of open field locomotor scores:effects of experience and teamwork on reliability. Multicenter Animal Spinal Cord Injury Study[J]. J Neurotrauma, 1996, 13(7):343-359.
22. Z'graggen WJ, Metz GA, Kartje GL, et al. Functional recovery and enhanced corticofugal plasticity after unilateral pyramidal tract lesion and blockade of myelin-associated neurite growth inhibitors in adult rats[J]. J Neurosci, 1998, 18(12):4744-4757.
23. Metz GA, Schwab ME, Welzl H. The effects of acute and chronic stress on motor and sensory performance in male Lewis rats[J]. Physiol Behav, 2001, 72(1/2):29-35.

Previous Article
Effects of EZH2-shRNA on Proliferation and Invasion of Human Glioma Cells
Next Article
Rat Model of Chronic Obstructive Pulmonary Disease Combined with Type 2 Diabetes Mellitus

West China Medical Journal

Effect of NEP1-40 Gene Modified Neural Stem Cell Transplantation on the Behavior Recovery of Rats after Spinal Cord Injury

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content