腺病毒介导的脑源性神经营养因子表达对大鼠损伤脊髓轴突的保护作用  

Protective effects of adenovirus-mediated expression of brain-derived neurotrophic factors on spinal axon after spinal cord injury in rats

作　　者：沈岳[1] 廖维宏[1] 王国强[1] 李芳[1] 伍亚民[1] 龙在云[1] 李应玉[1] 陈恒胜[1] 

机构地区：[1]第三军医大学附属大坪医院野战外科研究所创伤中心,重庆400042

出　　处：《第三军医大学学报》2000年第2期146-149,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金!(39670740)

摘　　要：目的 :研究腺病毒介导的脑源性神经营养因子(BDNF)基因在大鼠损伤脊髓内的表达规律 ,观察外源性BDNF对损伤脊髓的作用。方法 :利用激光束描记位移的重力打击装置 ,制备可靠的定量损伤模型 ;以直接注射法将带有BDNF基因表达框的复制缺陷重组腺病毒载体直接转移至大鼠损伤脊髓 ,用X Gal染色检测基因转移有效性 ,并观察损伤轴突计量形态学改变 ,损伤脊髓BDNFmRNA表达 ,BDNF和神经中丝(Neurofilament,NF)免疫组化活性的改变。结果 :腺病毒载体直接引入脊髓可有效地感染脊髓组织和表达报告基因。直接转染BDNF腺病毒载体可明显减少伤后两周轴突丧失 ;同时 ,NF免疫反应阳性的轴突较正常增多。结论 :实验性腺病毒介导的BDNF基因损伤脊髓转移是可行的。在基因直接转移局部外源性BDNF过表达对神经具有保护作用 ,可减少轴突丧失 ,促进神经元骨架蛋白的修复。这种双重作用可能在脊髓损伤治疗中有重要的价值。Objective: To investigate the effects of the expression of exogenous brain derived neurotrophic factors (BDNF) on injured spinal cord following in vivo adenoviral gene transfer to the cord. Methods: The model of quantitative injury was established in rats with the weight drop apparatus with a laser compression recorder. The replicate defect recombinant adenoviral vector was transferred into the injured spinal cord through direct injection. The gene transfer was verified with X Ga1 staining and the changes in morphology of the injured axon were quantitatively determined. Meanwhile, the expression of BDNF mRNA and immunohistochemical reactivity of BDNF and NF in the injured spinal cord were investigated. Results: The spinal cord was infected by injection of the adenoviral vector and expressed the report gene. The loss of axons reduced following the in vivo infection of adenoviral vector carrying exogenous BDNF gene after SCI, while more positive axons in immunohistochemical reaction than that of normal spinal cord were seen. Conclusion: In vivo transfer of adenoviral vector into the foci can provide protection to some extent to injured axons through increasing local expression of exogenous BDNF and facilitate repair of the cytoskeleton in the injured neurons. These effects are important in gene therapy of SCI.

关 键 词：疹髓损伤 腺病毒载体 基因治疗 大鼠 BDNF 

分 类 号：R651.2[医药卫生—外科学]