hBDNF-BMSCs局部移植对大鼠脊髓损伤的修复效果  

Effect of local transplantation of hBDNF-BMSCs on the repair of spinal cord injury in rats

作　　者：连学辉 肖红利 邓江[2] 韩子冀[1] 齐维林 LIAN Xuehui;XIAO Hongli;DENG Jiang;HAN Ziji;QI Weilin(Department of Orthopedics,Guiyang First People′s Hospital,Guiyang 550000,China;不详)

机构地区：[1]贵阳市第一人民医院骨科,贵阳550000 [2]遵义市第一人民医院骨科,贵州遵义563000

出　　处：《实用医学杂志》2021年第20期2590-2596,共7页The Journal of Practical Medicine

基　　金：国家自然科学基金委员会资助项目(编号:81660367);贵州省科学技术基金项目[编号:黔科合基础(2016)1420号]。

摘　　要：目的探讨hBDNF-BMSCs在大鼠脊髓内存活、分化、基因表达情况及其对大鼠脊髓损伤修复效果。方法提取并培养SD大鼠BMSCs,构建hBDNF-BMSCs。将SD大鼠随机分为假手术组、损伤对照组、空载体-BMSCs组、hBDNF-BMSCs组。采用改良Allen重物打击法建立大鼠脊髓损伤模型,1周后行hBDNF-BMSCs移植手术。采用BBB评分、皮层体感诱发电位评价各组大鼠治疗前后神经功能,并测定大鼠损伤脊髓节段中hBDNF的表达,观察大鼠损伤脊髓节段组织形态学变化,检测大鼠损伤脊髓节段内神经元核抗原(NeuN)、胶质纤维酸性蛋白(GFAP)、含H亚单位的磷酸化神经丝(p-NF-H)、神经生长相关蛋白(GAP-43)等表达。结果本研究成功提取BMSCs并构建hBDNF-BMSCs。hBDNF-BMSCs组大鼠神经功能改善明显优于空载体-BMSCs组及损伤对照组(P<0.05)。移植6周,hBDNF-BMSCs组大鼠下肢皮层体感诱发电位的P1波潜伏期明显缩短、P1-N1振幅最大。hBDNF-BMSCs组hBDNF表达水平1周时最高,6周时仍可见明显BDNF表达。hBDNF-BMSCs组和空载体-BMSCs组损伤区脊髓组织结构较完整,损伤对照组脊髓组织结构紊乱,并见胶质瘢痕形成。移植治疗6周后,hBDNF-BMSCs组与损伤对照组和空载体-BMSCs组相比,NeuN表达明显增多,GFAP表达明显减少,p-NF-H、GAP-43表达均明显增加并排列规整。结论hBDNF-BMSCs局部注射移植治疗SCI,可在局部持续稳定分泌hBDNF,保护神经元存活并促进移植的BMSCs转化为神经元样细胞,从而促进大鼠损伤脊髓损伤后的修复。Objective To investigate the survival,differentiation and gene expression of hBDNF-BMSCs in the spinal cord of rats and their repair effects on spinal cord injury(SCI)in rats.Methods The BMSCs of a SD rat were extracted and cultured to construct hBDNF-BMSCs.Another 96 SD rats were randomly divided into sham operation group,injury control group,empty vector-BMSCs group,hBDNF-BMSCs group.The rat SCI model was established by the modified Allen heavy object method,and hBDNF-BMSCs transplantation was performed one week later.BBB score and cortical somatosensory evoked potential(CSEP)were used to evaluate the nerve function of rats before and after treatment.The histomorphological changes of injured spinal cord segments in rats were observed.The expressions of NeuN,GFAP,p-NF-H and GAP-43 were detected by immunohistochemistry.Results The BMSCs was successfully extracted and the hBDNF-BMSCs was constructed.The neurological function in the hBDNF BMSCs group was significantly improved as compared with the vector-BMSCs group and the injury control group(P<0.05).After 6 weeks transplantation,the PI wave latency of the lower limb CSEP in the hBDNF-BMSCs group was significantly shortened,and the P1-N1 amplitude was the largest.The hBDNF expression in the hBDNF-BMSCs group was at the highest level in the first week,and even visible at 6 th week.In the hBDNF-BMSCs group and the empty carrier-BMSCs group,the spinal cord tissue structure in the injured area was relatively complete.The spinal cord tissue structure in the injured control group was disordered,and the glial scar formation was seen.As compared with the injury control group and the empty vector-BMSCs group,the NeuN expression was significantly increased,the GFAP expression was significantly reduced,and the p-NF-H,GAP-43 expressions were significantly increased and arranged regularly in the hBDNF-BMSCs group six weeks after transplantation.Conclusion Locally injected and transplanted hBDNF BMSCs can continuously and stably secrete hBDNF locally,protect the survival of ne

关 键 词：脊髓损伤 骨髓间充质干细胞 神经营养因子 基因治疗 神经修复 

分 类 号：R318[医药卫生—生物医学工程]