Intravenous transplantation of human umbilical cord blood mononuclear cells in an intracerebral hemorrhage rat model: microPET-CT evaluation and anti-inflammatory mechanisms

doi:10.3969/j.issn.2095-4344.0958

Abstract

Abstract:

BACKGROUND: Mononuclear cells isolated from umbilical cord blood (UCB-MNCs) are rich in cell types which are of great benefit in cell replacement therapy, gene therapy and tissue engineering.
OBJECTIVE: To evaluate the efficacy of human UCB-MNCs (HUCB-MNCs) in the treatment of intracerebral hemorrhage rats by microPET-CT and Longa scores, and to discuss the therapeutic mechanism.
METHODS: We made a rat intracerebral hemorrhage model by autologous blood injection. After modeling, rats were randomly divided into two groups: transplantation group and control group. The HUCB-MNCs were transplanted into the rats in the transplantation group via the tail vein. Control group received no treatment. Before and at 3, 7, 14, 21 days after transplantation, brain metabolic changes and neurological function scores of rats in the two groups were evaluated by microPET-CT and Longa scores. Levels of tumor necrosis factor-α and interleukin-6 in the rat serum of two groups were measured by ELISA method.
RESULTS AND CONCLUSION: (1) At 7, 14 and 21 days after HUCB-MNCs transplantation, the rates of brain metabolism in the hematoma center as well as around the hematoma in the transplantation group were significantly higher than that in the control group (P < 0.05), while the volume of hematoma and surrounding ischemic tissues as well as Longa scores were significantly reduced in the transplantation group as compared with the control group (P < 0.05). The levels of tumor necrosis factor-α and interleukin-6 in the serum in the transplantation group were significantly lower than those in the control group at 3, 7, 14 days after transplantation, and further lowered to the normal levels at 21 days after transplantation. These findings suggest that transplantation of HUCB-MNCs through the tail vein can promote the neurologic recovery of cerebral hemorrhage rats, and one of the therapeutic mechanisms may be related to the reduction of inflammatory factors and the inhibition of inflammatory response.

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: Tissue Engineering, Stem Cells, Tumor Necrosis Factor-alpha, Interleukin-6

CLC Number:

R743.34

Jin Na, Yao Xing-yu, Zhang Guo-hua, Yang Li-min, Li Jian-bo, Liu Jie, Wei Lin-huan, Duan Rui. Intravenous transplantation of human umbilical cord blood mononuclear cells in an intracerebral hemorrhage rat model: microPET-CT evaluation and anti-inflammatory mechanisms[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(25): 4014-4020.

Figures/Tables 2

2.1 实验动物数量分析纳入大鼠76只，其中10只作为ELISA分析时空白对照组大鼠，剩余66只大鼠造模，其中6只于造模过程中血液经针道返流或者死亡，实验过程中10只大鼠死亡，将死亡大鼠数据剔除，重新及时补充。实验过程中无大鼠感染。最终60只大鼠实验数据纳入结果分析。 2.2 人脐血单个核细胞活性将10 μL新鲜分离出的人脐血单个核细胞稀释至1 mL，加入体积分数0.5%的锥虫蓝3 μL摇匀，将10 μL上述稀释液滴加至细胞计数板上，置于光学显微镜下观察细胞活性并将细胞浓度调整为1×107 L-1(图1)。 2.3 大鼠神经行为学变化随机抽取造模大鼠，待麻醉清醒后行神经功能评分(图2A)，断头取脑解剖(图2B)，行microPET-CT扫描(图2C)验证造模成功。 2.4 移植组与对照组大鼠血肿中心代谢率(SUV%)比较两组大鼠不同时间点血肿中心代谢率均随时间变化呈逐渐增高的趋势(P < 0.05)；除移植前及移植后第3天，其他各个时间点两组大鼠血肿中心代谢率差异均有显著性意义 (P < 0.05)，且移植组大鼠血肿中心代谢率高于对照组(表1)。 2.5 移植组与对照组大鼠血肿周围代谢率比较两组大鼠不同时间点血肿周围代谢率均随时间推移逐渐增高(P < 0.05)；除移植前及移植后第3天，其他各时间点两组大鼠血肿周围代谢率差异均有显著性意义(P < 0.05)，且移植组大鼠血肿周围代谢率高于对照组(表2)。 2.6 移植组与对照组大鼠不同时间点血肿及周围缺血组织体积比较两组大鼠血肿及周围缺血组织体积随时间变化呈现逐渐减小的趋势(P < 0.05)；除移植前无显著差异外，其他各个时间点两组大鼠体积差异均有显著性意义(P < 0.05)，且移植组体积减小的趋势更明显(表3，图3)。 2.7 各组神经功能评分变化移植组与对照组大鼠不同时间点神经功能评分均随时间逐渐降低(P < 0.001)；除移植前及移植后第3天，其他各个时间点两组大鼠神经功能评分差异有显著性意义(P < 0.05)，且移植组评分较对照组低(图4)。 2.8 各组血清炎性因子变化各组大鼠不同时间点血清中肿瘤坏死因子α水平变化：空白组大鼠各个时间点血清中肿瘤坏死因子α均维持在较低水平。对照组与空白组比较：血清中肿瘤坏死因子α水平在大鼠脑出血后即升高，于第3天达高峰后逐渐下降，第21天降至与空白组水平，其他各时间点对照组大鼠血清中肿瘤坏死因子α水平均较空白组高(P < 0.05)；移植组与空白组相比：移植组大鼠移植前及移植后第3，7天血清中肿瘤坏死因子α水平较空白组明显升高(P < 0.05)，至第14，21天时降至与空白组相比无显著差异；移植组与对照组对比：两组大鼠移植前血清中肿瘤坏死因子α水平无显著差异，移植后的3，7，14 d，移植组大鼠血清中肿瘤坏死因子α水平均较对照组低(P < 0.05)，至第21天，两组大鼠血清中肿瘤坏死因子α水平无显著差异(表4)。各组大鼠不同时间点血清中白细胞介素6水平变化：空白组大鼠各个时间点血清中白细胞介素6水平均维持在一个稳定的低水平。对照组与空白组比较：大鼠血清中白细胞介素6水平在出血后即逐渐升高，于第7天达峰值后逐渐下降，于第21天降至与空白组无显著差异，其他各时间点血清中白细胞介素6水平均较空白组大鼠高(P < 0.05)；移植组与空白组相比：移植组大鼠移植前及移植后第3，7天血清中白细胞介素6水平较空白组明显升高(P < 0.05)，至第14，21天时降至与空白组相比无明显差异；移植组与对照组对比：两组大鼠移植前血清中白细胞介素6水平无显著差异，移植后第3，7，14天，移植组大鼠血清中白细胞介素6水平均较对照组低(P < 0.05)，第21天时两组大鼠血清中白细胞介素6差异无显著性意义(表5)。"

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