神经干细胞联合神经营养因子-3基因修饰的嗅鞘细胞移植对创伤性脑损伤大鼠神经功能的影响及机制  

Effect and mechanism of combined transplantation of neural stem cells and olfactory ensheathing cells modified by neurotrophin-3 gene on neural function of rats with traumatic brain injury

作　　者：张召宇 李梦欣 李慧堂 李海明 金保哲 ZHANG Zhaoyu;LI Mengxin;LI Huitang;LI Haiming;JIN Baozhe(Department of Neurosurgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;Henan Provincial Key Laboratory of Neural Repair,Weihui 453100,Henan Province,China)

机构地区：[1]新乡医学院第一附属医院神经外科,河南卫辉453100 [2]河南省神经修复重点实验室,河南卫辉453100

出　　处：《新乡医学院学报》2023年第6期501-506,共6页Journal of Xinxiang Medical University

基　　金：2018年河南省医学科技攻关计划省部共建项目(编号:SBGJ2018057)。

摘　　要：目的 探讨神经干细胞(NSCs)与神经营养因子-3(NT-3)基因修饰的嗅鞘细胞(OECs)联合移植对大鼠创伤性颅脑损伤(TBI)后神经功能的影响及机制。方法 将100只Sprague Dawley雄性大鼠按照随机数字表法分为假手术组、模型组、NSCs组、OECs组、联合治疗组,每组20只。模型组、NSCs组、OECs组、联合治疗组大鼠采用改良Feeney法构建TBI模型。造模1 d后,将不同的试剂注入各组大鼠损伤灶进行细胞移植,模型组大鼠注入10μL生理盐水,OECs组大鼠注入10μL NT-3基因修饰的OECs悬液,NSCs组大鼠注入10μL NSCs悬液,联合治疗组大鼠注入5μL NT-3基因修饰的OECs细胞悬液和5μL NSCs悬液,注射完成后继续常规饲养。分别于移植后1、7、14、28 d,各组随机取10只大鼠,采用改良神经功能缺损评分(mNSS)评估各组大鼠的神经功能缺损程度。移植后7 d,采用酶联免疫吸附试验法检测大鼠脑组织中白细胞介素(IL)-6、IL-1β及神经生长因子(NGF)水平。移植后28 d,采用苏木精-伊红染色法观察大鼠脑组织病理学改变,采用免疫组织化学染色法检测大鼠脑组织中NGF表达。结果 移植后1、7、14、28 d,模型组、NSCs组、OECs组、联合治疗组大鼠mNSS评分显著高于假手术组(P<0.05);移植后7、14 d,NSCs组、OECs组、联合治疗组大鼠mNSS评分显著低于模型组(P<0.05);移植后7、14 d,NSCs组、OECs组、联合治疗组大鼠mNSS评分比较差异无统计学意义(P>0.05);移植后28 d,NSCs组、OECs组、联合治疗组大鼠mNSS评分显著低于模型组,联合治疗组大鼠mNSS评分显著低于NSCs组和OECs组(P<0.05);移植后28 d,NSCs组与OECs组大鼠mNSS评分比较差异无统计学意义(P>0.05)。移植后28 d,假手术组大鼠脑组织结构完整,核仁明显,细胞排列有序;模型组大鼠损伤灶周围脑组织形态不规则,细胞肿胀,排列紊乱,可见坏死细胞及细胞核固缩,存在炎症细胞浸润;相较于模型组,NSCs组及OECs组大鼠脑组�Objective To explore the effect and mechanism of combined transplantation of neural stem cells(NSCs)and olfactory ensheathing cells(OECs)modified by neurotrophin-3(NT-3)gene on neurological function after traumatic brain injury(TBI)in rats.Methods A total of 100 male Sprague Dawley rats were randomly divided into sham operation group,model group,NSCs group,OECs group and combined treatment group using a random number table method,with 20 rats in each group.The rats in the model group,NSCs group,OECs group and combined treatment group were used to construct TBI models by using the improved Feeney method.After 1 d of modeling,different reagents were injected into the damaged lesion of rats in each group for cell transplantation,the rats in the model group injected with 10μL physiological saline,the rats in the OECs group injected with 10μL OECs modified by NT-3 gene suspension,the rats in the NSCs group rats injected with 10μL NSCs suspension,the rats in the combined treatment group injected with 5μL OECs modified by NT-3 gene cell suspension and 5μL NSCs suspension,routine feeding was performed after injection.On 1,7,14,28 d after transplantation,10 rats were randomly selected from each group and the degree of neurological impairment was evaluated by the modified neurological deficiency scale(mNSS).On 7 d after transplantation,the levels of interleukin(IL)-6,IL-1βand nerve growth factor(NGF)in brain tissue of rat were detected by enzyme-linked immunosorbent assay.On 28 d after transplantation,the histopathology changes in brain tissue of rats were detected by hematoxylin-eosin staining,and the expression of nerve growth factor(NGF)in brain tissue was detected by immunohistochemical staining.Results On 1,7,14,28 d after transplantation,the mNSS scores of rats in the model group,NSCs group,OECs group and combined treatment group were significantly higher than those in the sham operation group(P<0.05);on 7,14 d after transplantation,the mNSS scores of rats in the NSCs group,OECs group and combined treatment gro

关 键 词：神经生长因子 神经干细胞 嗅鞘细胞 创伤性脑损伤 

分 类 号：R651.15[医药卫生—外科学]