【摘要】 目的 :观察胶质细胞源性神经营养因子 (gliacellline derivedneurotrophicfactor ,GDNF)重组腺病毒 (Ad CMVgdnf)对培养的原代脊髓运动神经元的保护作用。 方法 :原代培养的脊髓运动神经元转染不同滴度Ad CMVgdnf不同时间后计数 ,并计算凋亡的运动神经元的百分率。 结果 :1 0 4 ～ 1 0 7pfu·ml 1 (pfu ,plaqueformunit)AdCMVgdnf的保护作用中 ,以 1 0 7pfu·ml 1 作用最显著 ,腺病毒的滴度过大或过小 ,保护作用都减弱 ;观察 1～ 9d不同转染时间的效果时 ,在第 3～ 9天均可观察到保护作用 ,以第 9天最明显 ,存活的运动神经元数比对照组增多2 80 % ,同时凋亡神经元的百分比较对照组减少 1 2 .9%。结论 :在一定的时间 (9天 )和合适的腺病毒滴度 (1 0 7pfu·ml 1 )条件下 ,腺病毒介导的GDNF对体外培养的脊髓运动神经元起到了保护作用。更多还原

【Abstract】 Objective: To observe the protective effects of recombinant glia cell line derived neurotrophic factor (GNDF) adenovirus (AdCMV gdnf ) on the cultured primary spinal motoneurons. Methods: After the transfection of AdCMV gdnf , immunocytochemistry of AChE was used to count the number of the cultured neurons, and the crystal violet was used to detect the percentage of the apoptosis neurons. Results: Considering the dose effect relationship, the protective effect was mostly significant at titer of 10 7 plaque form unit (pfu), the protective effects were weak when the titers of the virus were too high or too low as compared with 10 7 pfu·ml 1 ; Considering the time effect relationship, the recombinant AdCMV gdnf had significant protective effects after 3-9 days’ transfection with the most obvious effect on day 9. The survival rate of motoneuron increased by 280%. At the same time, the apoptotic percentage decreased by 12.8% . Conclusion: The recombinant GDNF adenovirus can protect the cultured primary motoneurons with appropriate titer and treatment time.更多还原