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中华细胞与干细胞杂志(电子版)

中华细胞与干细胞杂志(电子版) ›› 2017, Vol. 07 ›› Issue (02) : 71 -76. doi: 10.3877/cma.j.issn.2095-1221.2017.02.002

所属专题： 文献；

论著

脐带血单个核细胞对大鼠帕金森病的治疗效果观察
陈超1, 段婧1, 申爱芳2, 王伟1, 宋昊1, 芦现杰1, 王晓兵2, 韩忠朝3, 韩发彬1,†()   
  1. 1. 252000　聊城市人民医院　泰山医学院聊城临床学院干细胞与再生医学实验室
    2. 252000　聊城市人民医院　泰山医学院聊城临床学院妇产科
    3. 300020　天津，中国医学科学院，协和医科大学血液学研究所实验血液学国家重点实验室；300020　天津，国家干细胞工程技术研究中心
  • 收稿日期:2016-09-24 出版日期:2017-04-01
  • 通信作者: 韩发彬
  • 基金资助:
    山东省科技发展计划(2012 GSF11808); 山东省自然科学基金(ZR2015HL045)

Effects of human umbilical cord blood mononuclear cells in a rat model of Parkinson's disease

Chao Chen1, Jing Duan1, Aifang Shen2, Wei Wang1, Hao Song1, Xianjie Lu1, Xiaobing Wang2, Zhongchao Han3, Fabin Han1,†()   

  1. 1. Center for Stem Cells and Regenerative Medicine, the Liaocheng People's Hospital, Affiliated Liaocheng Hospital, Taishan Medical University, Liaocheng 252000, China
    2. Department of Gynecology and Obstetrics, the Liaocheng People's Hospital, Affiliated Liaocheng Hospital, Taishan Medical University, Liaocheng 252000, China
    3. the State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences, Peking Union of Medical College, Tianjin 300020, China; National Engineering Research Center of Cell Products, Tianjin 300020, China
  • Received:2016-09-24 Published:2017-04-01
  • Corresponding author: Fabin Han
  • About author:
    Corresponding author:Han Fabin, Email:
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引用本文：

陈超, 段婧, 申爱芳, 王伟, 宋昊, 芦现杰, 王晓兵, 韩忠朝, 韩发彬. 脐带血单个核细胞对大鼠帕金森病的治疗效果观察[J]. 中华细胞与干细胞杂志(电子版), 2017, 07(02): 71-76.

Chao Chen, Jing Duan, Aifang Shen, Wei Wang, Hao Song, Xianjie Lu, Xiaobing Wang, Zhongchao Han, Fabin Han. Effects of human umbilical cord blood mononuclear cells in a rat model of Parkinson's disease[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2017, 07(02): 71-76.

目的

探讨人体脐带血单个核细胞（HUCB-MNC）对大鼠帕金森病（PD）的治疗效果和作用机制。

方法

6-OHDA诱导产生大鼠PD模型，随机分为两组，一组移植从新生儿脐带血中分离HUCB-MNC细胞，另一组移植PBS作为对照，根据运动转圈数和转棒实验评估大鼠在细胞移植后不同时间的运动行为情况，移植16周后取出大脑进行冰冻切片，免疫荧光染色确定移植的细胞分化情况。动物行为采用Two-way ANOVA统计方法分析。

结果

转圈实验显示，PD大鼠移植HUCB-MNC细胞4、8、12、16周后，注射阿朴吗啡产生的转圈比率分别为0.529±0.121、0.457±0.096、0.369±0.089、0.264±0.091，而对照组4、8、12、16周转圈比率分别为0.843±0.158、0.812±0.156、0.736±0.103、0.714±0.160，与对照组相比，移植细胞组转圈数明显减少，差异有统计学意义（t = 4.377，5.561，6.835，6.071，P均< 0.01）；转棒实验显示，移植细胞大鼠在4、8、12、16周棒上持续时间分别为（46.333±2.082）s、（63.667±4.041）s、（76.667±7.638）s、（126±12.166）s，而对照组相应周数棒上持续时间分别为（29.250±5.560）s、（26.750±8.655）s、（34.250±10.905）s、（47.500±6.856）s，与对照组相比，移植细胞组棒上持续时间明显延长，差异有统计学意义（t = 4.966，6.189，5.707，10.990，P均< 0.01）。通过用人特异性细胞核抗体HNuc，神经细胞标志物Tuj1，和多巴胺神经元抗体TH染色，发现移植物HNuc、Tuj1、TH染色阳性。

结论

HUCB-MNC细胞可以分化为神经元和多巴胺神经元，减少PD大鼠阿朴吗啡诱导的转圈数，延长大鼠在转棒上的持续时间，改善PD大鼠模型的运动缺陷。

关键词: 脐血干细胞移植, 单核细胞, 帕金森病, 细胞分化, 多巴胺, 神经元
Objective

To explore whether human umbilical cord blood mononuclear cells (HUCB-MNC) transplantation could treat Parkinson's disease (PD) .

Methods

6-OHDA-lesioned PD rats were divided into two groups randomly: one group was treated with HUCB-MNCs, which were isolated from umbilical cord and the other group was treated with PBS as control. Apomorphine-induced rotation test was used to evaluate the motor behavior of the rats at different times after cell transplantation. Immunochemistry was conducted to confirm the differentiation of the grafted cells in the brain.

Results

The rates of apomorphine-induced rotation were 0.529±0.121, 0.457±0.096, 0.369±0.089, and 0.264±0.091 at 4, 8, 12, and 16 weeks after transplantation respectively, which were significantly lower than those of the control group (0.843±0.158, 0.812±0.156, 0.736±0.103, and 0.714±0.160 respectively) . And the duration time on rotation rod of the treatment group were (46.333±2.082) s, (63.667±4.041) s, (76.667±7.638) s, and (126±12.166) s respectively for each time points, while those of the control group were (29.250±5.560) s, (26.750±8.655) s, (34.250±10.905) s, and (47.500±6.856) respectively. Immunochemistry assay showed that the grafted HUCB-MNC expressed neuron maker Tuj1 and dopamine neuron maker TH.

Conclusion

Our study shows that transplanted HUCB-MNCs may differentiate into neurons and dopamine neurons and alleviate motor dysfunction of PD rats.

Key words: Cord blood stem cell transplantation, Monocytes, Parkinson disease, Cell differentiation, Dopamine, Neurons
图1 三种HUCB-MNC样本表面标记物平均表达率的流式检测
表1 两组PD大鼠移植后不同时间点转圈比率比较（ ± s）
分组 只数 4周 8周 12周 16周
对照组 7 0.843±0.158 0.812±0.156 0.736±0.103 0.714±0.160
移植组 7 0.529±0.121 0.457±0.096 0.369±0.089 0.264±0.091
t值 ? 4.377 5.561 6.835 6.071
P值 ? < 0.01 < 0.01 < 0.01 < 0.01
表2 两组PD大鼠移植后不同时间点棒上持续时间比较（s， ± s）
分组 只数 4周 8周 12周 16周
对照组 7 29.250±5.560 26.750±8.655 34.250±10.905 47.500± 6.856
移植组 7 46.333±2.082 63.667±4.041 76.667± 7.638 126.000±12.166
t值 ? 4.966 6.189 5.707 10.990
P值 ? < 0.01 < 0.01 < 0.01 < 0.01
图2 荧光显微镜下脐带血单个核细胞移植术后16周的PD大鼠脑内免疫组化分析（×100）
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