人脑源性神经营养因子基因修饰BMSCs联合EPO治疗大鼠急性脊髓损伤中的作用及机制

The effect and mechanism of h BDNF gene modified BMSCs combined with EPO in the treatment of acute spinal cord injury

目的观察人脑源性神经营养因子(h BDNF)基因修饰的间充质干细胞(BMSCs)在促红细胞生成素(EPO)治疗急性脊髓损伤中的治疗作用及其机制。方法通过病毒转染h BDNF基因到BMSCs后,将80只脊髓损伤模型SD大鼠,随机分为4组:NS组(在脊髓损伤前1 d经腹腔注射相同量的生理盐水)、EPO组(在脊髓损伤前按照5 000U/kg给予EPO)、联合组(脊髓损伤前给予5 000 U/kg EPO,损伤后在受损脊髓中点及受损中点上下5 mm中注入BDNF基因转染的BMSCs各5μl)和BDNF/BMSC组(脊髓损伤前腹腔给予等量生理盐水,在脊髓损伤后在脊髓中注入20μl BDNF基因转染BMSCs)。采用BBB法进行运动能力评分,PCR以及Western-blot法对脊髓BDNF基因及蛋白进行定量分析。结果较其他3组,联合组在脊髓损伤(SCI)早期与晚期对运动能力恢复效果最为明显,联合组在7 d、14 d、21d、28 d BDNF mRNA与蛋白表达水平均高于其他3组,差异有统计学意义(P<0.05)。结论 BDNF基因修饰的BMSC移植联合EPO治疗可促进大鼠脊髓损伤后神经功能的修复,这可能与促进脊髓损伤处BDNF的表达有关。

Objective To evaluate the effect and mechanism of hBDNF gene modified BMSCs combined with EPO in the treatment of a-cute spinal cord injury. Methods BDNF Gene was transfected by virus. Spinal cord injury rats were randomly divided into four groups：normal saline group,EPO group,BDNF/ BMSC group and combined group. The motor function was evaluated with BBB score and the RNA and protein of BDNF were measured by PCR and Western - blot. Results Combined group in SCI early and late on exercise ability to restore effect was most obvious,its mechanism may be related to SCI lesions,BDNF gene was associated with an increased content of protein expression. Conclusion The neurological function of rats with spinal cord can be obviously improved by autogeneicmesenchymal stem cells transplantation combined with EPO.