神经干细胞移植联合腹腔注射促红细胞生成素对横断性脊髓损伤大鼠神经轴突的修复作用

doi:10.13481/j.1671-587x.20150119

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (01): 99-104.doi: 10.13481/j.1671-587x.20150119

神经干细胞移植联合腹腔注射促红细胞生成素对横断性脊髓损伤大鼠神经轴突的修复作用

王喜良¹, 赵岩¹, 左媛², 肖宇龙¹, 霍洪军¹, 江建明³, 闫慧博⁴

1. 内蒙古医科大学第二附属医院胸腰椎脊柱外科, 内蒙古呼和浩特 010030;
2. 内蒙古医科大学第二附属医院检验科, 内蒙古呼和浩特 010030;
3. 南方医科大学附属南方医院脊柱外科, 广东广州 510515;
4. 南方医科大学第三附属医院脊柱外科, 广东广州 510630

收稿日期:2014-07-22 发布日期:2015-01-30
通讯作者: 赵岩,副主任医师,硕士研究生导师(Tel:0471-6351230,E-mail:nmgzy4568@126.com) E-mail:nmgzy4568@126.com
作者简介:王喜良(1988-),男,内蒙古自治区乌兰察布市人,在读医学硕士,主要从事脊柱脊髓损伤的基础和临床研究。
基金资助:
内蒙古自治区科技厅自然科学基金资助课题(2011MS1139)

Repaireffect of neural stem cells transplantation combined with intraperitoneal injection erythropoietin for axons in adult rats with transected spinal cord injury

WANG Xiliang¹, ZHAO Yan¹, ZUO Yuan², XIAO Yulong¹, HUO Hongjun¹, JIANG Jianming³, YAN Huibo⁴

1. Department of Thoracolumbar Spine Surgery, Second Affiliated Hospital, Inner Mongolia Medical University, Hohhot 010030, China;
2. Department of Clinical Laboratory, Second Affiliated Hospital, Inner Mongolia Medical University, Hohhot 010030, China;
3. Department of Orthopedic and Spinal Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China;
4. Department of Spinal Surgery, Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, China

Received:2014-07-22 Published:2015-01-30

摘要/Abstract

摘要：

目的: 探讨神经干细胞(NSCs)与促红细胞生成素(EPO)共同作用于横断性脊髓损伤大鼠后对损伤区轴突的修复作用,为临床治疗脊髓损伤提供理论依据。方法: 40只雌性成年Wistar大鼠,建立T10全横断大鼠脊髓损伤模型后,随机分为对照组、NSCs组、EPO组和联合治疗组,每组10只。术后8周采用BDA皮质脊髓束顺行追踪法和荧光金(FG)皮质脊髓束逆行追踪法评估损伤区脊髓神经轴突再生情况,同时分期采用实验性脊髓损伤运动功能BBB评分法评价大鼠后肢功能恢复情况。结果: BDA 免疫荧光染色和FG免疫荧光染色,联合治疗组可见大量被BDA-cy3红色荧光标记的再生轴突,其中部分再生轴突穿越损伤区到达远端;NSCs组仅见少量轴突再生,无神经轴突通过脊髓损伤区;EPO组偶见散在的神经纤维再生;对照组无明显的轴突再生。联合治疗组大脑皮质中可见少量被FG标记的椎体细胞及轴突发出金黄色荧光,其余3组大脑皮质中无FG标记细胞。大鼠后肢功能BBB评分,在术后1周及1周以后各时段,联合治疗组大鼠BBB评分均高于其他各组(P<0.05)。结论: 脊髓损伤后移植NSCs联合腹腔注射EPO可有效促进脊髓损伤区神经轴突的再生以及脊髓损伤大鼠后肢运动功能的恢复。

关键词: 神经干细胞, 促红细胞生成素, 干细胞移植, 全横断脊髓损伤, 轴突再生

Abstract:

Objective To investigate the repair effect of neural stem cells(NSCs) and erythropoietin (EPO) on the axonal regeneration of the rats with transected spinal cord injury in spinal cord injury area, and to provide theoretical basis for the clinical treatment of spinal cord injury. Methods Forty Wistar rats with T10 spinal cord transaction were randomly divided into control group, NSCs group, EPO group and NSCs+EPO group(n=10).BDA anterograde corticospinal cord neuronal tracing and fluoro-gold(FG) retrograde tracing were carried out at the 8th week after operation to observe the regeneration of nerve fibers.The BBB locomotion score was used to evaluate the restoration after operation in the rats with spinal cord transaction. Results The BDA immunofluorescence staining and FG immunofluorescence staining results showed that a large amount of regenerated axons were observed in NSCs+EPO group, which were labeled by BDA-cy3 red fluorescence, and some regenerated axons got through the injuried area.While in NSCs group, only a small amount of regenerated axons were observed after spinal cord injury, and no regenerated axons got through the injuried area.In EPO group, only sporadically regenerated nerve fibers were observed occasionally, while in control group, no axonal regeneration was observed.In NSCs+EPO group, a small amount of cones and axons, which were labeled by FG fluorescence, emitted golden yellow fluorescence, and no cells labeled by FG was observed in other groups.The BBB scores in NSCs+EPO group 7 d after operation and at different time later were higher than those in other groups(P<0.05). Conclusion Transplantated NSCs and intraperitoneal injection of EPO could promote axon regeneration in spinal cord injury zone and hind limb motor function recovery of the rats with spinal cord injury.

Key words: neural stem cells, erythropoietin, stem cell transplantation, transected spinal cord injury, axonal regeneration

中图分类号:

R681.54

王喜良, 赵岩, 左媛, 肖宇龙, 霍洪军, 江建明, 闫慧博. 神经干细胞移植联合腹腔注射促红细胞生成素对横断性脊髓损伤大鼠神经轴突的修复作用[J]. 吉林大学学报(医学版), 2015, 41(01): 99-104.

WANG Xiliang, ZHAO Yan, ZUO Yuan, XIAO Yulong, HUO Hongjun, JIANG Jianming, YAN Huibo. Repaireffect of neural stem cells transplantation combined with intraperitoneal injection erythropoietin for axons in adult rats with transected spinal cord injury[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 99-104.

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神经干细胞移植联合腹腔注射促红细胞生成素对横断性脊髓损伤大鼠神经轴突的修复作用

Repaireffect of neural stem cells transplantation combined with intraperitoneal injection erythropoietin for axons in adult rats with transected spinal cord injury

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