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.
出处
《中国脊柱脊髓杂志》
CAS
CSCD
北大核心
2012年第3期272-277,共6页
Chinese Journal of Spine and Spinal Cord
关键词
脊髓损伤
脑红蛋白
转基因
丙二醛
一氧化氮
Spinal cord injury
Neuroglobin
Gene transfer
Malondialdehyde
Nitric oxide
