慢病毒介导的脑红蛋白体内基因转染对兔损伤脊髓的作用  

The protection of lentivirus-mediated neuroglobin gene transfer in vivo on injured spinal cord in rabbits,an experimental study

作　　者：陈宣维[1] 林文平[2] 林建华[1] 

机构地区：[1]福建医科大学附属第一医院骨脊柱外科,福州市350005 [2]福建医科大学附属第二医院骨科,泉州市362000

出　　处：《中国脊柱脊髓杂志》2012年第3期272-277,共6页Chinese Journal of Spine and Spinal Cord

摘　　要：目的:观察慢病毒介导脑红蛋白(Ngb)体内基因转染兔损伤的脊髓组织后对后肢运动功能的影响,探讨其作用机制。方法:用球囊压迫法制成兔脊髓损伤(SCI)模型96只,随机分为对照组(A组)、生理盐水组(B组)、空载体组(C组)和Ngb慢病毒组(D组),每组动物24只,A组SCI后无治疗;B组SCI后向脊髓内注射生理盐水;C组SCI后向脊髓内注射空病毒;D组SCI后向脊髓内注射Ngb重组慢病毒。各组分别在1、3、7、14、21d采用BBB运动功能评分系统检测兔后肢运动功能情况;观察损伤脊髓组织内标记荧光的表达;Real-timePCR和Western blot检测Ngb mRNA及其相应蛋白的表达情况,生化方法检测丙二醛(MDA)和一氧化氮(NO)水平。结果:损伤后14d和21d,D组BBB评分明显高于其他3组(P<0.05),但A、B及C组之间比较无差异(P>0.05);C组和D组兔损伤部位脊髓组织均有GFP表达的绿色荧光信号;损伤后7d、14d和21d,D组的Ngb表达与其他3组比较明显增强(P<0.05);损伤后7d、14d和21d,D组损伤脊髓组织中MDA、NO含量明显低于其他3组(P<0.05)。结论:慢病毒介导脑红蛋白(Ngb)体内基因转染可使Ngb高表达,可能是通过减轻继发性SCI,从而促进SCI后后肢运动功能的恢复。Objectives： To investigate the possible protective mechanism of lentivirus-mediated neuroglobin gene transfer in vivo on injured spinal cord（SCI） in rabbits.Methods： Ninety-six New Zealand white rabbits were randomly divided into four groups（n=24） according to intervention after SCI： no intervention（A）;normal saline group（B）;LV-GFP group（C）;and LV-Ngb group（D）.SCI was introduced by epidural balloon compression.24 hours after compression,T9 laminectomy was performed,then normal saline,LV-GFP or LV-Ngb was injected in situ respectively.Motor function of hind limbs after injection was evaluated by Basso-Beattie-Bresnahan（BBB） score,the expressions of green fluorescence in injured spinal cord were observed by fluorescence microscope,the expressions of Ngb mRNA and protein in injured spinal cord were detected by real-time PCR and Western blot,and the levels of malondialdehyde（MDA） and nitric oxide（NO） were also tested.Results： Significant functional improvement was observed in group D compared with group A,B and C at day 14 and 21.The expressions of green fluorescence in injured spinal cord were observed in group C and D.The Ngb expression in group D increased significantly compared with the other 3 groups at day 7,14 and 21.The MDA and NO levels in group D were lower than the other 3 groups（P〈0.05） at day 7,14 and 21.Conclsions： Lentivirus-mediated neuroglobin gene transfer in vivo may induce high expression of Ngb,which may be resulted from decreasing secondary spinal cord injury,thus promoting hind limbs motorfunction.

关 键 词：脊髓损伤 脑红蛋白 转基因 丙二醛 一氧化氮 

分 类 号：R683.2[医药卫生—骨科学] Q786[医药卫生—外科学]