Adipose mesenchymal stem cells for treatment of traumatic brain injury

doi:10.3969/j.issn.2095-4344.2017.01.013

Abstract

Abstract:

BACKGROUND: Clinical and animal studies have confirmed that transplanted mesenchymal stem cells can migrate to the area of brain injury, and exert a certain therapeutic effect on traumatic brain injury.
OBJECTIVE: To study the therapeutic effect of adipose mesenchymal stem cells on the local damage induced by traumatic brain injury in rats.
METHODS: According to the literatures, the brain injury model of Sprague-Dawley rats was made by brain freezing injury. After successful modeling, the rats were randomly divided into three groups: model rats in experimental group were administrated with transplantation of adipose mesenchymal stem cells via the tail vein at 2 days after modeling; model rats in control group given the same amount of normal saline; and normal rats in normal group given no treatment. Morris water maze test was used to evaluate the recovery of neurological function in rats. The isolated and cultured adipose mesenchymal stem cells were observed under an inverted microscope. The distribution of these cells in the injured brain and the levels of brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) were detected by immunohistochemistry method.
RESULTS AND CONCLUSION: Morris water maze test results showed that compared with the control group, the average escape latency of rats decreased rapidly in the experimental group, and the difference was statistically significant (P < 0.05), while the escape latency of rats in the experimental group was gradually close to that in the control group. Immunohistochemical findings showed that Brdu-labeled adipose mesenchymal stem cells were accumulated and distributed unevenly in the injured cerebral cortex. In the control group, there was no BrdU immunofluorescence staining in the rat brain tissues. Western blot test results showed that: compared with the control group, the BDNF and GDNF levels in the hippocampus of rats were significantly higher in the experimental group (P < 0.05), but compared with the normal group, the BDNF level in the hippocampus of rats was significantly increased, and the GDNF level in the cortex decreased significantly in the control group (P < 0.05). These findings indicate that adipose mesenchymal stem cells can migrate to the damaged area of rats, and promote the secretion of BDNF and GDNF in the injured brain, which may be one of the mechanisms by which adipose mesenchymal stem cell transplantation improves neurological functional recovery of rats.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Mesenchymal Stem Cells, Stem Cell Transplantation, Brain-Derived Neurotrophic Factor, Glial Cell Line-Derived Neurotrophic Factor

CLC Number:

R394.2

Zhu Jun-qing, Hong Jun, Cui Jian-zhong, Wang Kai-jie. Adipose mesenchymal stem cells for treatment of traumatic brain injury[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(1): 71-76.

Figures/Tables 2

2.1 实验动物数量分析实验中所用到的50只SD大鼠全部进入结果分析，无脱失。 2.2 体外培养的脂肪间充质干细胞检测结果在体外培养的SD大鼠腹腔肠系脂肪间充质干细胞经流式细胞仪检测，所分离出的细胞培养整合达90%左右时，脂肪间充质干细胞呈扁圆形和纺锤形，符合脂肪间充质干细胞的形态，具有较好的自我更新能力(图1)。 2.3 SD大鼠外伤性脑损伤建模结果实验采用脑冷冻伤制作大鼠外伤性脑损伤模型。各组SD大鼠脑冷冻伤建模后没有出现死亡现象，对大鼠的活动没有显著影响，可以正常进行Morris水迷宫测试。苏木精-伊红染色检测显示，建模后的大鼠大脑皮质损伤区域出现局限性损伤，各大鼠脑损伤区域基本一致，可见附近细胞核的碎裂、坏死、凋亡，表明本实验所用模型建模成功(图2)。 2.4 各组大鼠Morris水迷宫测试结果各组大鼠测量的第1，2，3，4，5天的平均潜伏期，采用单个重复测量因素的方差分析，5次重复测量的数据间存在高度的相关性。脂肪干细胞移植后Morris水迷宫测试潜伏期第1，2，3，4，5天后，实验组与对照组比，实验组SD大鼠平均逃避潜伏期迅速下降，差异有显著性意义(P < 0.05)，而同时实验组大鼠逃避潜伏期越来越接近于正常组大鼠，结果见表1。各组潜伏期指标总体而言不同，可以看出，实验组的潜伏期低于对照组，而略高于正常组的潜伏期，3组SD大鼠的潜伏期随测量时间逐渐缩短。 2.5 干细胞在大鼠脑损伤区域中的分布 BrdU标记的第4代脂肪间充质干细胞经SD大鼠尾静脉移植后，免疫组织化学后置于倒置显微镜观察在大鼠脑细胞内的分布结果显示，移植的经Brdu标记的脂肪间充质干细胞大量聚集在损伤的大脑皮质，呈不均匀分布。而对照组大鼠脑组织内未见BrdU免疫荧光显色。说明脂肪间充质干细胞经大鼠尾静脉移植后能通过大鼠血脑屏障迁移至外伤性损伤的大鼠的受损区域，进一步在大脑组织中分散(图3)。 2.6 大鼠脑组织中脑源性神经营养因子和胶质细胞源性神经营养因子含量检测结果与对照组比，实验组经脂肪干细胞静脉移植后SD大鼠脑源性神经营养因子、胶质细胞源性神经营养因子的含量明显高于对照组(P < 0.05)。而与正常组SD大鼠比较，对照组大鼠脑源性神经营养因子含量升高，而皮质周围的胶质细胞源性神经营养因子含量却明显下降(表2)。单向方差分析显示，各组SD大鼠脑组织损伤部位或附近的脑源性神经营养因子与胶质细胞源性神经营养因子的相对表达量有明显的差异性，同时组间比较差异有显著性意义(P < 0.05)。"

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