中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (6): 835-839.doi: 10.12307/2023.231

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

临床级人脐带间充质干细胞对创伤性脑损伤大鼠神经功能的改善作用

崔连旭1，江文康2，陆大鸿1，许峻荣3，刘小翠3，王丙云2   

  1. 1佛山市第一人民医院微创及脊髓神经外科，广东省佛山市   528000；2佛山科学技术学院生命科学与工程学院，广东省佛山市   528225；3广东唯泰生物科技有限公司，广东省佛山市   528251
  • 收稿日期:2022-01-06 接受日期:2022-03-18 出版日期:2023-02-28 发布日期:2022-08-10
  • 通讯作者: 崔连旭，硕士，副教授，佛山市第一人民医院微创及脊髓神经外科，广东省佛山市 528000 王丙云，博士，教授，佛山科学技术学院生命科学与工程学院，广东省佛山市 528225
  • 作者简介:崔连旭，男，1970年生，辽宁省沈阳市人，汉族，硕士，副主任医师，副教授，主要从事创伤性脑损伤研究。 江文康，男，1996年生，广东省肇庆市人，汉族，在读硕士，主要从事动物干细胞方面的研究。
  • 基金资助:
    佛山市登峰计划课题：重点项目(2019)24，项目负责人：崔连旭

Clinical-grade human umbilical cord mesenchymal stem cells affect the improvement of neurological function in rats with traumatic brain injury

Cui Lianxu1, Jiang Wenkang2, Lu Dahong1, Xu Junrong3, Liu Xiaocui3, Wang Bingyun2   

  1. 1Department of Minimally Invasive and Spinal Neurosurgery, Foshan First People’s Hospital, Foshan 528000, Guangdong Province, China; 2School of Life Science and Engineering, Foshan University of Science and Technology, Foshan 528225, Guangdong Province, China; 3Guangdong VitaLife Biotechnology Co., Ltd., Foshan 528251, Guangdong Province, China
  • Received:2022-01-06 Accepted:2022-03-18 Online:2023-02-28 Published:2022-08-10
  • Contact: Cui Lianxu, Master, Associate professor, Department of Minimally Invasive and Spinal Neurosurgery, Foshan First People’s Hospital, Foshan 528000, Guangdong Province, China Wang Bingyun, MD, Professor, School of Life Science and Engineering, Foshan University of Science and Technology, Foshan 528225, Guangdong Province, China
  • About author:Cui Lianxu, Master, Associate chief physician, Associate professor, Department of Minimally Invasive and Spinal Neurosurgery, Foshan First People’s Hospital, Foshan 528000, Guangdong Province, China Jiang Wenkang, Master candidate, School of Life Science and Engineering, Foshan University of Science and Technology, Foshan 528225, Guangdong Province, China Cui Lianxu and Jiang Wenkang contributed equally to this article.
  • Supported by:
    Key Project of Foshan Dengfeng Plan Subject, No. (2019) 24 (to CLX) 

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摘要：

文题释义：
临床级脐带间充质干细胞：在成熟的干细胞制备工艺和完善的质量控制体系下的GMP标准的细胞工厂生产，经中国食品药品检定院质量认定并获得《检验报告》，可用于临床运用的间充质干细胞。
创伤性脑损伤：是临床上常发生的一种因头部被撞击、突然加速或者突然减速等机械力造成的脑部损伤疾病，导致患者的运动能力和学习记忆能力障碍，严重影响患者的生活质量。

背景：创伤性脑损伤是世界范围内发生病率最高、最严重的神经系统疾病之一，暂无有效治疗方法，有研究表明间充质干细胞对创伤性脑损伤有一定的修复作用。
目的：观察临床级人脐带间充质干细胞对创伤性脑损伤大鼠神经功能的改善作用。
方法：将45只雌性SD大鼠随机分为3组，分别为假手术组(n=15)、模型组(n=15)和干细胞组(n=15)，除假手术组外，其余两组采用改良的Feeney自由落体方法建立创伤性脑损伤模型，造模24 h后，模型组脑定位注射20 μL生理盐水，干细胞组注射20 μL临床级人脐带间充质干细胞悬液，共1×106个细胞，细胞移植后1，3，7，14，21 d采用改良神经功能缺损评分评估大鼠的神经功能运动；移植后21 d，采用RT-qPCR检测Bcl-2/Bax mRNA比值，RT-qPCR及免疫荧光法检测胶质细胞标记物GFAP和IBA1的表达，ELISA法检测血浆中肿瘤坏死因子α、白细胞介素6和白细胞介素10水平，苏木精-伊红染色观察脑组织结构。
结果与结论：①与模型组相比，干细胞组改良神经功能缺损评分明显降低，大鼠的运动功能明显改善；②与模型组相比，干细胞组脑组织Bcl-2/Bax mRNA比值显著升高，胶质细胞标记物GFAP和IBA1 mRNA的表达显著降低，血浆肿瘤坏死因子α、白细胞介素6和白细胞介素10水平差异不显著；③苏木精-伊红染色观察干细胞组大鼠脑组织缺损、水肿和神经元凋亡等情况均有不同程度的改善；④结果表明，在创伤性脑损伤后移植临床级人脐带间充质干细胞可促进神经功能恢复，减少神经元细胞凋亡，抑制胶质细胞过度活化。

https://orcid.org/0000-0002-4935-0656(崔连旭) 

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

关键词: 脐带间充质干细胞, 创伤性脑损伤, 神经功能恢复, 胶质细胞, 凋亡

Abstract: BACKGROUND: Traumatic brain injury is one of the most serious neurological diseases with the highest incidence in the world, and there is no effective therapy at present. Studies have shown that mesenchymal stem cells have a certain repair effect on traumatic brain injury. 
OBJECTIVE: To investigate the effects of clinical-grade umbilical cord mesenchymal stem cells on the improvement of neurological function in rats with traumatic brain injury. 
METHODS:  Forty-five rats were divided into three groups, namely sham operation group (n=15), model group (n=15) and stem cell group (n=15). Except the sham operation group, the modified Feeney free fall method was used to establish the traumatic brain injury model in the model and stem cell groups. At 24 hours after model establishment, the model group was implanted with 20 μL normal saline; and the stem cell group was implanted with 20 μL human umbilical cord mesenchymal stem cell suspension, a total of 1×106 cells. At 1, 3, 7, 14, and 21 days after cell transplantation, the neural function of the rats was evaluated by modified neurological severity scores. At 21 days after injection, the ratio of Bcl-2/Bax mRNA was detected by RT-qPCR. The expression levels of gliocyte markers GFAP and IBA1 were detected by RT-qPCR and immunofluorescence method. At 21 days after injection, plasma levels of tumor necrosis factor alpha, interleukin 6 and interleukin 10 were determined by ELISA. Hematoxylin-eosin staining was used to observe the brain tissue structure. 
RESULTS AND CONCLUSION: (1) Compared with the model group, modified neurological severity score was significantly lower and the motor function was significantly improved in the stem cell group. (2) Compared with the model group, Bcl-2/Bax mRNA ratio was significantly increased in brain tissue, the expression of GFAP and IBA1 was significantly decreased in the stem cell group, while the expression of tumor necrosis factor alpha, interleukin 6 and interleukin 10 in plasma was not significantly different. (3) Hematoxylin-eosin staining showed that the defect area and edema of brain tissue and neuronal apoptosis were improved in different degrees in the stem cell group. (4) The results show that clinical-grade human umbilical cord mesenchymal stem cells can promote neurological function recovery, reduce neuronal apoptosis and inhibit glial overactivation after traumatic brain injury.

Key words: umbilical cord mesenchymal stem cell, traumatic brain injury, neurological function recovery, glial cell, apoptosis

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