中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2012, Vol. 41 ›› Issue (4): 307–310.

• 基础医学 • 上一篇    下一篇

Ephrin-B3沉默对大鼠脊髓损伤后功能修复及Gap-43表达的影响

杜鹏,吕刚,智晓东   

  1. (辽宁医学院附属第一医院骨科,辽宁 锦州 121001)
  • 收稿日期:2012-09-24 修回日期:2012-09-24 出版日期:2012-04-20 发布日期:2012-09-26

Effects of Silence Ephrin-B3 on Functional Recovery and Expression of Growth Associated Protein-43 after the Spinal Cord Injury in Rats

DU Peng,LǚGang,ZHI Xiao-dong   

  1. (Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical College, Jinzhou 121001, China)
  • Received:2012-09-24 Revised:2012-09-24 Online:2012-04-20 Published:2012-09-26

摘要: 目的 观察Ephrin-B3基因沉默后对脊髓损伤(SCI)功能恢复的影响。方法 构建3组针对Ephrin-B3基因的siRNA干扰质粒(用于实验组:A,B,C组)及1组乱序siRNA干扰质粒(用于对照组D组),体外克隆并提取质粒后包装慢病毒颗粒。半横切法制作成年大鼠SCI模型,将病毒颗粒注入SCI处。分别在注入1,2,4,8周后,采用BBB评分法比较大鼠肢体功能恢复情况;采用Real-time PCR法检测伤后各组脊髓组织Ephrin-B3 mRNA表达情况;采用免疫组化法观察脊髓组织Gap-43的表达变化。结果 Ephrin-B3 mRNA在正常大鼠脊髓内呈相对低表达,D组表达水平呈上升趋势,实验组大鼠出现Ephrin-B3沉默效果,B组尤为明显。转染后1~4周,B组大鼠BBB评分呈持续升高趋势,高于其他3组,D组明显低于实验组,4周时各组恢复效果都进入平台期。免疫组化结果显示,B组脊髓组织内Gap-43表达最明显,表达趋势从转染后1周持续至4周,8周时有所下降,而D组呈持续性弱表达。结论 沉默Ephrin-B3能够促进大鼠SCI早期功能修复并上调脊髓组织内Gap-43的表达。

关键词: 脊髓损伤, 大鼠, 功能修复, Ephrin-B3, Gap-43

Abstract: Abstract Objective To observe the effects of silence Ephrin-B3 on functional recovery after spinal cord injury (SCI) in rats. Methods Three groups of siRNA interference plasmid for Ephrin-B3 (used in three experimental group: A, B,C) and one group of siRNA interference plasmid (used in control group: D) were constructed, cloned in vitro, extracted and packed into slow virus particles. Adult rat SCI model was established by half-crosscut and injected with slow virus particles. One, two, four and eight weeks after the injection respectively, BBB evaluation method was used to compare limb functional recovery in four groups; Real-time PCR method was used to detect the expression of Ephrin-B3 mRNA in spinal tissues after injury; immunohistochemical method was used to observe the expression of Gap-43 changes in spinal tissues. Results The expression of Ephrin-B3 was relatively low in normal rats, showed clear ascendant trend in group D and silenced in experimental groups, especially in group B. During the first four weeks after the transfection, BBB score of group B kept rising, higher than those in other three groups, while that in group D was significantly lower than the experimental groups. At the end of the fourth week after the transfection, the recovery of all the groups entered the platform stage. Immunohistochemical results showed Gap-43 was expressed most significantly in group B and the expression continued to increase during the first four weeks after the transfection and decreased at the eighth week. Conclusion Silence Ephrin-B3 can promote the functional recovery after SCI in rats at early stage, and raise the expression of Gap-43 in spinal tissues.

Key words: spinal cord injury, sats, functional recovery , Ephrin-B3 , GAP-43

中图分类号: 

  • R651.2 文献标志码 A
[1] Chadi G, Andrade MS, Leme RJ, et al. Experimental models of partial lesion of rat spinal cord to investigate neurodegeneration, glial activation,and behavior impairments[J]. Int J Neurosci,2001,111(3-4):137-165.
[2] Basso DM,Beattie MS,Bresnahan JC. A sensitive and reliable lo-comotor rating scale for open field testing in rats[J]. J Neurotrauma,1995,12(1) : 1-21.
[3] 赵玉斌,张积仁.乳腺癌EphA2,EphrinAl;ERa,ERp和PR的表达及RNAi沉默EphA2基因的实验研究[D].南方医科大学,2010.
[4] Shao Z, Browning JL, Lee X, et al. TAJ/TROY, an orphan TNF receptor family member, binds Nogo-66 receptorl and regulates axonal regeneration[J]. Neuron, 2005, 45(3): 353-359.
[5] Benson MD, RomeroMI, Lush ME, et al. Ephrin-B3 is a myelin-based inhibitor of neurite out-growth [J]. PNAS, 2005, 102(30): 10694-10699.
[6] 肖琴,陈娟,吴艳秋. 脑室周围白质软化新生大鼠脑组织 ephrin-b3 表达及神经细胞凋亡[J]. 中国当代儿科学杂志,2007.9(4):321-323.
[7] Yiu G, He Z. Signalingmechanisms of themyelin inhibitors ofax-on regeneration[J]. Curr Opin Neurobio J, 2003, 13(5): 545-551.
[8] Yokoyama N, Romero MI, Cowan CA, et al. Forward signaling mediated by ephrin-B3 prevents contralateral corticospinal axons from recrossing the spinal cordmidline [J]. Neuron, 2001,29(1):85-97.
[9] Goldshmit Y, Galea MP, Wise G, Bartlett PF&Turnley AM. Axonal Regeneration and Lack ofAstrocytic Gliosis in EphA4-Deficient Mice. J. Neurosci,2004,24(45):10064-10073.
[10] 张在强,曹世俭,王拥军.实验性糖尿病大鼠坐骨神经损伤后神经再生微环境变化[J] .首都医科大学学报,2009,30(5): 664-670.
[11] Davies SFA, FitchMT, Memberg SP, et al. Regeneration of adult axons in white matter tracts of the central nervous system[J]. Nature, 1997, 390(6661):680-683.
[1] 吴越, 周祖华, 闻庆平, 苗壮. 葛根素对大鼠背根神经节神经病理性疼痛的镇痛作用[J]. 中国医科大学学报, 2018, 47(9): 803-806.
[2] 解丽丽, 吕洪涛, 许瑞雪. 磷酸化P38在神经病理性疼痛大鼠背根神经节中的作用机制[J]. 中国医科大学学报, 2018, 47(4): 308-311.
[3] 邢海会, 丁晓慧, 解辉, 王忠华, 谢菊华, 陈丰洋, 罗崟洲, 周胜男. Toll样受体4对大鼠脑缺血再灌注损伤的作用[J]. 中国医科大学学报, 2018, 47(3): 206-211.
[4] 徐春卓, 辛颖. 宫内发育迟缓对大鼠胰腺中Ngn3表达及内分泌细胞发育的影响[J]. 中国医科大学学报, 2018, 47(10): 891-894,899.
[5] 赵彦楠,刘洪祥,马晓欣. 晚期糖基化终产物受体在多囊卵巢综合征大鼠模型中的表达[J]. 中国医科大学学报, 2017, 46(9): 769-772.
[6] 刘晓颖 ,姚俊洁 ,刘文. 白藜芦醇对链脲霉素诱导的糖尿病大鼠神经病理性疼痛的作用[J]. 中国医科大学学报, 2017, 46(7): 591-594.
[7] 赵彦楠 ,刘洪祥,霍佳宁,马晓欣. 晚期糖基化终末产物在多囊卵巢综合征大鼠模型中的表达[J]. 中国医科大学学报, 2017, 46(5): 388-391.
[8] 桑亮 ,王学梅 ,许东阳 ,桑力轩 ,商功群 ,袁胜美 ,屠树星. 超声量化指标预测大鼠肝纤维化严重程度[J]. 中国医科大学学报, 2017, 46(5): 429-433.
[9] 季相禄,田峰,王斌,肖万安. 葛根素治疗急性脊髓缺血再灌注损伤大鼠的神经保护机制[J]. 中国医科大学学报, 2017, 46(4): 313-316.
[10] 张瑛瑛,李季泓. 甘草对高脂血症大鼠模型洛伐他汀药代动力学的影响[J]. 中国医科大学学报, 2017, 46(4): 342-344.
[11] 李晓倩,包娜仁,张再莉,马虹. 大麻素受体 1 对坐骨神经结扎大鼠神经病理性疼痛的影响及机制[J]. 中国医科大学学报, 2017, 46(3): 205-209.
[12] 孙德日 ,胡慧媛 ,朱悦. 改良脊髓半横断损伤大鼠模型的建立及评价[J]. 中国医科大学学报, 2017, 46(3): 223-226.
[13] 刘瑶,赵晓云,韩颖,赵雅宁,李建民,李佳宁. 脑缺血后处理对脑缺血再灌注大鼠Notch1信号通路及 学习记忆功能的影响[J]. 中国医科大学学报, 2017, 46(11): 984-989.
[14] 冯瑞,赵玫. 钙蛋白酶及其抑制剂对心肌肥厚模型大鼠心肌肥厚作用的研究[J]. 中国医科大学学报, 2016, 45(9): 769-772.
[15] 蔡洪梅 ,王维 ,马頔 ,王文娟 ,易添 ,周晓红. 经皮电刺激联合减重跑台训练对完全性脊髓损伤大鼠胶质源性 神经营养因子及 RhoA 表达的影响[J]. 中国医科大学学报, 2016, 45(9): 838-842.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!

中国医科大学学报版权所有©2018

未经允许,严禁擅自转载本站图文资料

地址:中国 沈阳市沈北新区蒲河路77号 110122

辽ICP备05014850

JOURNAL OF CHINA MEDICAL UNIVERSITY

ADDRESS: NO.77 PUHE ROAD

SHENYANG NORTH NEW AREA, SHENYANG

LIAONING PROVINCE, P.R. CHINA