摘要
目的研究神经营养因子-3（NT-3）基因修饰的嗅鞘细胞移植对自身免疫性脑脊髓炎（EAE）的髓鞘及轴突的修复作用。方法用逆转录病毒介导神经营养因子-3（NT-3）基因转染嗅鞘细胞（OEC），将其移植入EAE侧脑室，荧光标记后观察其在体内的迁徙、分布特点；运用皮质体感诱发电位监测（CSEP）、超微结构观察、功能评分；RT—PCR方法检测NT-3mRNA转录水平，并与对照组、OEC移植组相比较等方法对髓鞘及轴突修复进行评价。结果（1）OEC-NT-3在EAE体内存活，可广泛迁徙至病灶远端且至少可以持续存活4周。（2）超微结构发现转基因治疗组4周后，轴索结构完整，周围髓鞘板层结构清楚，明显优于另外二组。（3）4周后，转基因组CSEP潜伏期明显缩短，波幅增加，明显优于其他二组，差异有统计学意义（P〈0．05）。（4）转基因组NT-3mRNA转录水平为（212．32±16．14）×10^-2，明显高于OEC组（1．23±0．13）×10^-2及对照组（1．98±0．19）×10^-2，差异有统计学意义（P〈0．01）。结论NT-3基因修饰的OEC在持续表达神经营养因子，且可促进EAE的神经修复。
Objective To investigate the therapeutic effect of neurotrophin-3 （NT-3） modified olfactory ensheathing cell （OEC） upon experimental allergic encephalomyelitis （EAE）. Methods OEC- NT-3 gene engineering cell, constructed by neurotrophin-3 transinfecting OEC inducted by retrovirus, was transplanted into lateral ventricle. The migration and distribution were observed and compared with control group and OEC transplantation group. Then myelin repairing and axon regeneration were evaluated from cortical somatosensory evoked potential （CSEP）, function score and ultrastructural morphology. Results （ 1 ） OEC-NT-3 could survive, migrate within axons and spread diffusely away from the focus at Day 28 post- transplantation; （2）as compared with other two groups, more nerve fibers, better myelin repair and more distinct myelin structure were observed in the transgene group; （3）as compared with other two groups, the latent time was obviously shortened and the amplitude higher in the transplantation group （P 〈0. 05） ; （4） the transcription level of NT-3mRNA in the transgene group was significantly higher than the OEC group and the contrast group （212.32 ±16. 14） ×10^-2 vs （1.98 ±0.19）×10^-2, （1.23 ±0.13） ×10^-2（P 〈 0. 01 ）. Conclusion OEC-NT-3 cell expresses NT-3 stably and effectively in EAE. It may contribute to the repairing of myelin and the regeneration of axon.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2009年第29期2063-2067,共5页
National Medical Journal of China
基金
国家自然科学基金（30770751）
山东省卫生厅青年基金（2007QZ002）
关键词
嗅鞘细胞
神经营养因子-3
基因
自身免疫性脑脊髓炎
Olfactory ensheathing cell
Neurotrophin-3
Gene
Experimental allergic encephalomyelitis
