NGF过表达质粒修饰BMSCs移植促进大鼠急性脊髓损伤修复

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Abstract Objective To investigate the effect of bone marrow mesenchymal stem cell transplantation transfected with nerve growth factor (NGF) gene overexpression vector on the repair of acute spinal cord injury in rats.Methods Human bone marrow mesenchymal stem cells (BMSCs) were extracted and the surface proteins of BMSCs were detected by flow cytometry.NGF overexpression vector plasmid was constructed by Cas-9 technology.BMSCs were transfected by lentivirus technology.The animal models of acute spinal cord injury in SD rats wereestablished by clamp method.SCI was constructed according to different treatment schemes.The models were divided into three groups: model group，BMSCs group and overexpression plasmid group.There were5models in each group.The NGF expression and nerve repair were detected by RT-PCR，Western blot and the immunofluorescence.Results The expression of CD44 and CD29 was 60.2% and 58.3% in the second generation BMSCs，while the expression of CD34 and CD45 was 3.4% and 2.6% in the second generation BMSCs by flow cytometry.The transfection efficiency of lentivirus was over 90%.The BBB score of the model group was the lowest one day after operation，and that of the overexpression plasmid group was higher than that of the model group and BMSCs group (P＜0.05).Compared with model group and BMSCs group，the vacancy rate of hind limbs in overexpression plasmid group was significantly higher (P＜0.05).RTPCR results showed that the relative expression level of NGF in overexpression plasmid group was significantly higher than that in model group and BMSCs group (P＜0.05).The result of Western blot was the same as that of RT-PCR.Immunofluorescence results showed that NGF could co-stain with NeuN in the overexpressed plasmid group，and the number of neurons increased.The degree of spinal cord repair in overexpression plasmid group was better than that in BMSCs group and model group.It was found that the fracture spacing was obviously reduced and the repair of the broken end was better.Conclusion NGF overexpression plasmid can promote BMSCs to differentiate into neurons and promote the functional recovery of SCI animal model.

Key words： nerve growth factor bone marrow mesenchymal stem cells spinal cord injury lentivirus transfection

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	Shi Yong
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Shi Yong,Xia Xiaoyan. Transplantation of BMSCs Modified with NGF Overexpression Plasmid Promotes Repair of Acute Spinal Cord Injury in Rats[J]. sygkzz, 2020, 26(8): 707-715.

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http://www.sygkzz.com/EN/ OR http://www.sygkzz.com/EN/Y2020/V26/I8/707

［1］朱双龙，段会全，刘英富，等.柴胡皂苷a对大鼠急性脊髓损伤的神经保护作用与机制研究［J］.中国修复重建外科杂志，2017，31(7)：825-829.
［2］Assinck P，Duncan GJ，Hilton BJ，et al.Cell transplantation therapy for spinal cord injury［J］.Nature Neuroscience，2017，20(5)：637-647.
［3］Zhao H，Chen S，Gao K，et al.Resveratrol protects against spinal cord injury by activating autophagy and inhibiting apoptosis mediated by the SIRT1/AMPK signaling pathway［J］.Neuroscience，2017(348)：241-251.
［4］Burke D，Fullen BM，Stokes D，et al.Neuropathic pain prevalence following spinal cord injury：A systematic review and meta-analysis［J］.Eur J Pain，2017，21(1)：29-44.
［5］Ruzicka J，Machova-Urdzikova L，Gillick J，et al.A comparative study of three different types of stem cells for treatment of rat spinal cord injury［J］.Cell Transplant，2017，26(4)：585-603.
［6］Liu L，Sun T，Xin F，et al.Nerve growth factor protects against alcohol-induced neurotoxicity in PC12 cells via PI3K/Akt/mTOR pathway［J］.Alcohol Alcohol，2017，52(1)：12-18.
［7］Lee H，Shin CH，Kim HR，et al.MicroRNA-296-5p promotes invasiveness through downregulation of nerve growth factor receptor and caspase-8［J］.Mol Cells，2017，40(4)：254-261.
［8］杨进顺，吕浩然，廖壮文，等.大鼠急性脊髓损伤动物模型的建立及影像学分析［J］.广东医学，2011，32(5)：559-561.
［9］Nejati-koshki K，Mortazavi Y，Pilehvar-soltanahmadi Y，et al.An update on application of nanotechnology and stem cells in spinal cord injury regeneration［J］.Biomed Pharmacother，2017(90)：85-92.
［10］Seo DK，Kim JH，Min J，et al.Enhanced axonal regeneration by transplanted Wnt3a-secreting human mesenchymal stem cells in a rat model of spinal cord injury［J］.Acta Neurochir(Wien)，2017，159(5)：947-957.
［11］Zhao H，Cheng L，Du X，et al.Transplantation of cerebral dopamine neurotrophic factor transducted BMSCs in contusion spinal cord injury of rats：promotion of nerve regeneration by alleviating neuroinflammation［J］.Mol Neurobiol，2016，53(1)：187-199.
［12］陈乃耀，申娜，贺靖澜，等.人脐血间充质干细胞治疗鼠创伤性脑损伤后BDNF的变化［J］.山东大学学报：医学版，2013，51(1)：22-26.
［13］杨方方，刘萍，程静，等.慢病毒介导shRNA沉默大鼠肝BRL-3A细胞PDZK1对胆管侧膜Bsep表达的影响［J］.安徽医科大学学报，2019，11(9)：1338-1342.
［14］缪晓刚，阿布都艾尼·热吾提，王利，等.基质金属蛋白酶-11在骨肉瘤中的表达及生物学功能研究［J］.实用骨科杂志，2018，24(11)：46-49.