移植NRG1转染的雪旺细胞对脊髓损伤大鼠mTOR信号通路的影响  

Effect of transplantation of Schwann cells transfected with NRG1 on p-mTOR signaling pathway in rats with spinal cord injury

作　　者：尤福利 杨明超[1] 李毅[1] 周鑫 吴立莹 王岩峰[1] YOU Fu-li;YANG Ming-chao;LI Yi;ZHOU Xin;WU Li-ying;WANG Yan-feng(Department of Orthopedics, the First Affiliated Hospital of China Medical University, Shenyang, 110001, China)

机构地区：[1]中国医科大学附属第一医院骨科,辽宁沈阳110001

出　　处：《解剖科学进展》2018年第3期314-317,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金青年基金(81100924);辽宁省自然科学基金优秀人才培育(2014021016)

摘　　要：目的探讨移植NRG1转染的雪旺细胞对脊髓损伤大鼠mTOR信号通路的影响。方法体外分离培养雪旺细胞,免疫荧光鉴定;将pcDNA3.1-NRG1重组质粒通过Fugen6转染雪旺细胞,用免疫荧光法检测细胞转染率。参照Nystrom's压迫方法制作大鼠脊髓压迫损伤模型,于伤后第7天移植转基因雪旺细胞,细胞移植7周后结束实验,每周对实验动物进行BBB评分。实验分为假手术组、脊髓损伤组与转基因雪旺细胞移植组。Western blot方法检测各组大鼠脊髓损伤部位p-mTOR蛋白的表达。结果成功实现激活态雪旺细胞体外分离、培养和鉴定,并成功转染雪旺细胞,细胞转染率为62.25%。与脊髓损伤组相比,在细胞移植后的第3周开始,转基因雪旺细胞移植组大鼠的BBB评分显著升高(P<0.05)。与假手术组比较,脊髓损伤组与转基因雪旺细胞移植组大鼠脊髓损伤部位p-mTOR的蛋白表达水平显著降低(P<0.05),但转基因雪旺细胞移植组比脊髓损伤组显著升高(P<0.05)。结论移植NRG1转染的雪旺细胞能够有效促进脊髓损伤大鼠神经功能的恢复,其机制可能与激活mTOR信号通路有关。Objective To investigate the effect of transplantation of Schwann cells transfected with NRG1 on mTOR signaling pathway in rats with spinal cord injury. Methods Schwann cells were isolated and cultured in vitro and identified by immunofluorescence. The recombinant plasmid pc DNA3.1-NRG1 was transfected into Schwann cells by Fugen 6, and the transfection rate was detected by immunofluorescence. Spinal cord injury rat model was made with the Nystrom＇s method. Transgenic Schwann cells were transplanted on the 7 th day after injury, and the experiment was completed 7 weeks after cell transplantation.The BBB score of the experimental animals was evaluated weekly. The experiment was divided into the sham operation group, the spinal cord injury group and the transgenic Schwann cell transplantation group. The expression of p-mTOR in the site of spinal cord injury was detected by Western blot. Results The activated Schwann cells were successfully isolated, cultured and identified in vitro. The transfection rate of the cells was 62.25%. Compared with spinal cord injury group, BBB score of the rats in transgenic Schwann cells transplantation group was increased significantly the third weeks after cell transplantation（P〈0.05）. The expression level of p-mTOR was significantly decreased in spinal cord injury group than in sham group, but significantly increased in genetically modified Schwann cells transplantation than in spinal cord injury group（P〈0.05）. Conclusion Transplantation of Schwann cells transfected with NRG1 could effectively promote the recovery of neural function in rats with spinal cord injury, which might be related to the activation of mTOR signaling pathway.

关 键 词：脊髓损伤 NRG1 雪旺细胞 转染 MTOR 大鼠 

分 类 号：R681.5[医药卫生—骨科学]