神经营养素3及神经干细胞联合移植治疗实验性大鼠缺氧缺血性脑损伤  

Combined transplantation of neurotrophin-3 and neural stem cells in treatment of hypoxic-ischemic brain injury in rats

作　　者：王丽敏[1] 王海燕[1] 杨自金[2] 朱晓峰[3] 原德新[4] 

机构地区：[1]佳木斯大学附属第一医院儿科,154003 [2]山东省菏泽医学专科学校儿科教研室 [3]佳木斯大学神经科学研究所 [4]佳木斯大学组胚教研室,154003

出　　处：《中华儿科杂志》2007年第3期212-216,共5页Chinese Journal of Pediatrics

摘　　要：目的研究神经营养素3(NT-3)及神经干细胞联合移植对缺氧缺血性脑损伤(HIBD)大鼠神经功能恢复及神经干细胞分化为神经元的比例的影响,并探讨其可能机制。方法取新出生的24 h 内的 wistar 大鼠,从海马中分离神经细胞,进行培养、鉴定。用出生7 d 的 wistar 新生大鼠制作缺氧缺血性脑损伤的动物模型,模型成功后7 d 进行移植。将实验大鼠随机分为5组:正常组、模型组、假移植组、神经干细胞移植组、NT-3及神经干细胞联合移植组(简称联合移植组),每组12只。移植部位为损伤同侧侧脑室。移植后4周进行功能实验,取脑组织进行免疫组化及免疫荧光检查。结果从新生鼠海马中成功培养出神经干细胞,培养条件下呈悬浮状态生长,形成神经球,绝大多数的细胞表达神经干细胞的标志物神经巢蛋白(nestin)。说明培养的细胞大部分为神经干细胞,细胞纯度达90%以上。Y 迷宫实验结果:神经干细胞移植组平均(163±11.6)次学会,记忆的平均正确率为(50±13.3)%;神经干细胞(NSC)+NT-3联合移植组平均(117.27±11.04)次学会,记忆的正确率平均(63±11.2)%。握持牵引实验结果:神经干细胞移植组平均(30.1±11.8)s,右下肢能放置的占40%;NSC+NT-3联合移植组平均(40.64±10.6)s,右下肢能放置的占72.73%。斜坡实验结果:神经干细胞移植组平均(20.3±8.25);NSC+NT-3联合移植组平均(12.9±5.15)s。说明接受 NT-3及神经干细胞移植组大鼠的学习能力、记忆能力及肢体功能与单纯神经干细胞移植组相比有明显提高,差异具有统计学意义(P<0.05)。其神经干细胞分化成神经元的比例(50%)与单纯神经干细胞移植组(30%)比有明显增多。结论 NT-3与神经干细胞联合移植能提高缺氧缺血性脑损伤大鼠的学习、记忆能力和肢体功能,并能提高神经干细胞向神经元转化的比率,NT-3和神经干细胞联合移植较单独神经干细胞移植对缺氧缺血性脑损�Objective To evaluate the efficacy of combined transplantation of neurotrophin-3 and neural stem ceils in treatment of hypoxic-ischemic brain injury in rats, and study the possible mechanism. Methods Neural stem cells （ NSCs ） isolated from hippocampi of newborn wistar rats were cultured and identified. The hypoxic-ischemic brain injury models established with 7-day-old wistar rats, and neural stem cells or others were transplantated into the ipsilateral ventricles 7 days later. The 7-day-old rats were randomized into 5 groups： normal group, model group, sham-transplantation group, NSCs transplantation group, and NT-3 combined with NSCs transplantation group. There were 12 rats per group. The functional test and immunohistochemistry were examined 4 weeks later. Results The neural stem cells from new-born rats＇ hippocampi were successfully cultured. It was found that they formed typical neurospheres in suspension, and the majorities of cells expressed nestin, which was the marker for neural stem cells. The rats from combined transplantation group performed significantly better in the ability to study and in memory and the hmb function than the rats from NSCs transplantation group （P 〈 0. 05 ）. The rate of neural stem cells differentiating into neurons from combined transplantation group was higher, too （50% vs. 30% ）. Conclusion Combined NT-3 and NSCs transplantation could improve the abihty to study, memory and the hmb function of rats after hypoxic-ischemic brain injury, and improve the rate of NSCs differentiating neurons. Combined NT-3 and NSCs transplantation had better effects on hypoxic-ischemic rats than transplantation of NSCs alone.

关 键 词：神经营养素3 神经干细胞 移植 缺氧缺血性脑损伤 

分 类 号：R722.1[医药卫生—儿科]