AxCA-脑源性神经营养因子转染大鼠损伤坐骨神经后基因表达的检测  

Expression of BDNF gene after gene transfer of AxCA-BDNF to the sciatic nerve for peripheral nerve regeneration in rats

作　　者：李培建[1] 李兵仓[2] 

机构地区：[1]北京军区总医院神经外科,100700 [2]第三军医大学大坪医院野战外科研究所六室

出　　处：《中华实验外科杂志》2006年第12期1436-1438,I0018,共4页Chinese Journal of Experimental Surgery

基　　金：国家重点基础研究发展规划专项资助项目(G1999054206)

摘　　要：目的观察携带小鼠脑源性神经营养因子(BDNF)cDNA表达片段的非依赖辅助复制缺陷型重组腺病毒载体(AxCA)-BDNF转染大鼠坐骨神经后基因表达情况。方法成年大鼠随机分成A组(坐骨神经缺损+硅胶管+AxCA—BDNF原液8μl);B组(坐骨神经缺损+硅胶管+BD—NF溶液8μl);C组(坐骨神经缺损+硅胶管+空白病毒稀释液8μl)三组。应用原位杂交和免疫组织化学检测方法,从BDNF mRNA和蛋白水平,进行坐骨神经损伤后BDNF基因表达的定性和半定量分析测定。结果伤后腺病毒介导的BDNF基因转移组,在3、7、14 d、1个月,4个时相点,近、远端神经干和脊髓(L3-6)中BDNF mRNA水平均远远高于其他两组,BDNF的水平也远远高于作为对照的单纯硅胶管套接组。结论通过腺病毒介导转染的BDNF基因在大鼠坐骨神经SCs内得到了有效表达,并通过轴突逆行转运到了相应的脊髓神经元。Objective To observe the expression of BDNF gene after microinjected adenovirusmediated gene transfer of BDNF （AxCA-BDNF） to the sciatic nerve for peripheral nerve regeneration. Methods Based on silicone tube graft as a support to bridge adult rat sciatic nerve gaps, Wistar rats were divided into 3 group receiving microinjection of recombinant adenovirus vector of BDNF （AxCA-BDNF）, BDNF and simple injection of virus buffer to the sciatic nerve respectively. By using the methods of in situ hybridization and immunocytochemistry, the BDNF gene expression was detected. Results It was certified the BDNF gene was delivered into the Schwann cells and expressed efficiently. The expression of the BDNF gene lasted for 1 month, within the month nerve regeneration was accomplished in the rats. Conclnsion The exogenous BDNF gene and its expression proteins were up-taken to the spinal cord motoneurons through retrograde axonal transport.

关 键 词：坐骨神经 腺病毒 基因表达 免疫组织化学 原位分子杂交 

分 类 号：R651.3[医药卫生—外科学]