解剖学报

解剖学报 ›› 2021, Vol. 52 ›› Issue (2): 205-209.doi: 10.16098/j.issn.0529-1356.2021.02.007

• 神经生物学 • 上一篇    下一篇

壳聚糖包载人脐带间充质干细胞对创伤性脑损伤大鼠的治疗作用及机制

张宇1 薛茜2* 宋笑雨1 尹卫东3 邹玉安2   

  1. 1. 河北北方学院研究生学院， 河北 张家口 075000;2. 河北北方学院附属第一医院神经内科，河北 张家口 075000; 3. 河北北方学院继续教育学院， 河北 张家口 075000
  • 收稿日期:2019-09-02 修回日期:2019-10-20 出版日期:2021-04-06 发布日期:2021-04-06
  • 通讯作者: 薛茜 E-mail:starxincheng@163.com
  • 基金资助:
    河北省2015年度科学研究重点课题

Therapeutic effect and mechanism of hydrogel-encapsulated human umbilical cord mesenchymal stem cells on traumatic brain injury in rats

ZHANG Yu1  XUE Qian2* SONG Xiao-yu1  YIN Wei-dong3  ZOU Yu-an2    

  1. 1. Graduate College of Hebei North University, Hebei Zhangjiakou 075000, China; 2. Department of Neurology, the First Affiliated Hospital of Hebei North University, Hebei Zhangjiakou 075000, China; 3. Continuing Education College of Hebei North University, Hebei Zhangjiakou 075000, China
  • Received:2019-09-02 Revised:2019-10-20 Online:2021-04-06 Published:2021-04-06
  • Contact: XUE Qian E-mail:starxincheng@163.com

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摘要：

目的 探讨壳聚糖包载人脐带间充质干细胞（hUC-MSCs）对创伤性脑损伤大鼠的治疗效果及相关机制。  方法 构建40只创伤性脑损伤SD大鼠模型，根据干预方案的不同，分为对照组(n=10)和壳聚糖组(n=10)、干细胞组(n=10)和联合治疗组(n=10)，采用mNSS量表评价大鼠运动功能，Morris水迷宫法评价学习记忆能力，利用Western blotting法检测神经元特异性烯醇化酶(NSE)、神经元特异性核蛋白(NeuN)、微管相关蛋白 2(MAP2)、脑源性神经营养因子（BDNF）和B淋巴细胞瘤-2基因(Bcl-2)蛋白表达，比较几种治疗方式的差异。   结果  术后第1天、4天和8天，各组大鼠的mNSS评分基本一致（P＞0.05）。在治疗过程中，对照组与壳聚糖组的mNSS评分差异无显著性（P＞0.05）。在第15天、22天、29天和36天，与对照组相比，联合治疗组mNSS评分下降幅度最明显，干细胞组次之（P<0.05）。水迷宫实验结果显示，随着治疗时间的延长，各组大鼠的逃避潜伏时间均逐渐降低。与对照组相比，联合治疗组逃避潜伏时间下降幅度最明显，干细胞组次之（P<0.05）；同时干细胞组与联合治疗组大鼠所在象限停留时间显著长于对照组及壳聚糖组（P＜0.05）。免疫荧光结果显示，在蓝色背景下，治疗组大鼠脑组织内呈红色荧光表现的β-Ⅲ tubulin阳性的细胞数量显著高于对照组。与对照组相比，壳聚糖组的NSE、NEUN、MAP2、BDNF和Bcl-2蛋白表达量无明显变化（P＞0.05），干细胞组及联合治疗组相关蛋白表达量明显升高。两两比较后发现，联合治疗组相关蛋白的表达量最高，干细胞组次之，对照组及壳聚糖组最低，差异均具有统计学意义（P＜0.05）。   结论 与单纯干细胞移植相比，壳聚糖包载hUC-MSCs移植可通过促进受损部位细胞再生，有效地改善创伤性脑损伤大鼠的运动及学习记忆能力。

关键词: 间充质干细胞, 脑损伤, 水凝胶, 免疫印迹法, 大鼠

Abstract:

Objective To investigate the therapeutic effect of hydroge-encapsulated human umbilical cord mesenchymal stem cells (hUC-MSCs) on traumatic brain injury in rats and its related mechanism.   Methods SD rat models of traumatic brain injury were constructed, which were divided into control group, chitosan group, stem cell group and combined treatment group.    Results During the treatment, there was no significant difference in mNSS score between the control group and the chitosan group (P>0.05). On the 15th, 22nd, 29th and 36th day, the mNSS score of the combined treatment group decreased most significantly than that of the control group, followed by the stem cell group(P<0.05). Compared with the control group, the escape latency of the combined treatment group decreased most significantly, followed by the stem cell group (P<0.05). The number of beta-Ⅲ tubulin-positive cells in the brain tissues of rats in the treatment group was significantly higher than that in the control group. Compared with the control group, there was no significant change in the expression of neuron-specific enolase (NSE), neuron-specific nuclear protein (NeuN), microtubule associated protein 2 (MAP2), brain-derived neurotrophic factor (BDNF) and B cell lymphoma-2 (Bcl-2) in the chitosan group (P>0.05), and the expression of related proteins in the stem cell group and the combined treatment group increased significantly. The expression level of the related protein in the combined treatment group was the highest, followed by stem cell group, the lowest in the control group and chitosan group (P<0.05).    Conclusion Compared with stem cell transplantation alone, hydrogel-encapsulated hUC-MSCs transplantation can improve the motor and learning and memory abilities of rats with traumatic brain injury more effectively.

Key words: Mesenchymal stem cell, Brain injury, Hydrogel, Western blotting, Rat

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