论文部分内容阅读
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 study the expression of adenovirus-mediated brain-derived neurotrophic factor (BDNF) gene in rat spinal cord injury and to observe the effect of exogenous BDNF on injured spinal cord. Methods: Reliable quantitative damage model was prepared by laser beam tracing gravity displacement device. The replication-deficient recombinant adenovirus vector with BDNF gene expression cassette was directly transferred to the injured spinal cord by direct injection method and detected by X-Gal staining The gene transfer efficiency was observed. The morphological changes of injured axons, the expressions of BDNF mRNA, the expression of BDNF and the activity of neurofilament (NF) in injured spinal cord were observed. Results: The direct introduction of adenovirus vector into spinal cord effectively infected spinal cord tissue and expressed the reporter gene. Direct transfection of BDNF adenovirus vector significantly reduced axonal loss two weeks after injury; at the same time, NF-positive axons increased more than normal. Conclusion: Experimental adenovirus-mediated BDNF gene injury in spinal cord is feasible. In the direct gene transfer of local exogenous BDNF over-expression on the nerve has a protective effect, can reduce the loss of axons and promote repair of neuronal scaffold proteins. This dual role may have important value in the treatment of spinal cord injury.