中国组织工程研究 ›› 2020, Vol. 24 ›› Issue (19): 2972-2977.doi: 10.3969/j.issn.2095-4344.2057

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

人脐血间充质干细胞移植脑梗死大鼠神经功能的恢复及脑组织端粒酶反转录酶表达

余  恒,周  涛   

  1. 襄阳市中心医院,湖北省文理学院附属医院北院区神经内科,湖北省襄阳市  441003
  • 收稿日期:2019-07-25 修回日期:2019-08-01 接受日期:2019-09-17 出版日期:2020-07-08 发布日期:2020-04-08
  • 通讯作者: 周涛,主治医师,襄阳市中心医院,湖北省文理学院附属医院北院区神经内科,湖北省襄阳市 441003
  • 作者简介:余恒,男,1978年生,湖北省丹江口市人,汉族,2002年武汉大学毕业,主治医师,主要从事癫痫、痴呆方面的研究。
  • 基金资助:
    湖北省自然基金项目(2013CCV064)

Effects of umbilical cord blood-derived mesenchymal stem cell transplantation on recovery of nerve function and telomerase reverse transcriptase expression in brain tissue of cerebral infarction rats 

Yu Heng, Zhou Tao   

  1. Department of Neurology, Xiangyang Central Hospital, North Branch of Affiliated Hospital of Hubei University of Arts and Science, Xiangyang 441003, Hubei Province, China
  • Received:2019-07-25 Revised:2019-08-01 Accepted:2019-09-17 Online:2020-07-08 Published:2020-04-08
  • Contact: Zhou Tao, Attending physician, Department of Neurology, Xiangyang Central Hospital, North Branch of Affiliated Hospital of Hubei University of Arts and Science, Xiangyang 441003, Hubei Province, China
  • About author:Yu Heng, Attending physician, Department of Neurology, Xiangyang Central Hospital, North Branch of Affiliated Hospital of Hubei University of Arts and Science, Xiangyang 441003, Hubei Province, China
  • Supported by:
    the Natural Science Foundation of Hubei Province, No. 2013CCV064

摘要:

文题释义:

人脐血来源间充质干细胞的优势:脐血从孕妇分娩后的胎盘、脐带残端收集,对于产妇和新生儿均没有任何痛苦和不良影响,也不会涉及社会、伦理及法律方面的争论;脐血受胎盘屏障的保护,其成分被病毒、细菌污染的概率低。

端粒酶:细胞中负责端粒延长的一种酶,是基本的核蛋白反转录酶,在保持端粒稳定、基因组完整、细胞长期活性和潜在的继续增殖能力等方面有重要作用。

背景:脐血间充质干细胞可以经体循环有效穿透室管膜进入脑组织,迁移至脑损伤区域并存活分化为神经细胞,作为替代细胞来源用于中枢神经系统疾病的治疗。

目的:探讨人脐血间充质干细胞移植对脑梗死大鼠神经功能恢复及脑组织端粒酶反转录酶表达的影响。

方法:选取130只大鼠为研究对象,随机分为对照组(30只)、模型组(30只)、脐血间充质干细胞1组(35只)、脐血间充质干细胞2组(35只);对照组不做任何处理,其余各组采用颈内动脉线栓法制作脑梗死大鼠模型,脐血间充质干细胞1,2组于造模成功后1,4 d尾静脉移植2×106个脐血间充质干细胞;移植后24 h检测大鼠血清中活性氧和超氧化物歧化酶水平,移植后第7,14天进行Y-迷宫实验和神经功能评分,采用TUNEL法检测病灶中心神经细胞凋亡率,Western blot法检测梗死灶周围Caspase-3、Bax、Bcl-2、端粒酶反转录酶蛋白表达,RT-PCR法检测梗死灶周围端粒酶反转录酶的mRNA表达。

结果与结论:①与对照组相比,模型组大鼠Y-迷宫实验错误次数、神经功能评分、细胞凋亡指数、Caspase-3、Bax蛋白表达、活性氧水平均显著升高(P < 0.05),Bcl-2、端粒酶反转录酶蛋白以及mRNA表达、超氧化物歧化酶水平均显著降低(P < 0.05);②与模型组相比,脐血间充质干细胞1组和脐血间充质干细胞2组大鼠Y-迷宫实验错误次数、神经功能评分、细胞凋亡指数、Caspase-3、Bax蛋白表达、活性氧水平均显著降低(P < 0.05),Bcl-2、端粒酶反转录酶蛋白以及mRNA表达、超氧化物歧化酶水平均显著升高(P < 0.05);③与脐血间充质干细胞2组相比,脐血间充质干细胞1组大鼠Y-迷宫实验错误次数、神经功能评分、细胞凋亡指数、Caspase-3、Bax蛋白表达、活性氧水平均显著降低(P < 0.05),Bcl-2、端粒酶反转录酶蛋白以及mRNA表达、超氧化物歧化酶水平均显著升高(P < 0.05);④结果表明,脐血间充质干细胞移植可以有效促进脑梗死大鼠神经功能恢复,能够提高脑组织端粒酶反转录酶表达,移植时间越早效果越显著。

ORCID: 0000-0002-4902-1553(余恒)

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: 脑梗死, 脐血间充质干细胞, 神经功能, 端粒酶反转录酶, 细胞凋亡

Abstract:

BACKGROUND: Umbilical cord blood-derived mesenchymal stem cells can penetrate the ependyma into the brain tissue via the systemic circulation, migrate into the injury area and differentiate into neurons. They are as substitute cells applied in the treatment of central nervous system diseases

OBJECTIVE: To explore the effects of umbilical cord blood-derived mesenchymal stem cell transplantation on the functional recovery of nerve and expression of telomerase reverse transcriptase in the brain tissue of cerebral infarction rats.

METHODS: One hundred and thirty rats were enrolled and were randomly divided into control group (n=30), model group (n=30), umbilical cord blood-derived mesenchymal stem cell 1 group (n=35) and umbilical cord blood-derived mesenchymal stem cell 2 group (n=35). The control group was not given any treatment. The rat models of cerebral infarction were made by internal carotid artery suture method in the other groups. At 1 and 4 days after successful modeling, 2×106 umbilical cord blood-derived mesenchymal stem cells were transplanted into tail vein in the umbilical cord blood-derived mesenchymal stem cell 1 and 2 groups. The levels of reactive oxygen species and superoxide dismutase were detected at 24 hours after transplantation. The nerve function score was detected by Y-maze test at 7 and 14 days after transplantation. The neuronal apoptosis in the center of lesions was detected by TUNEL assay. The expression levels of Caspase-3, Bax, Bcl-2 and telomerase reverse transcriptase proteins around the infarct lesion were detected by western blot assay. The mRNA expression of telomerase reverse transcriptase around infarct lesions was detected by RT-PCR.

RESULTS AND CONCLUSION: Compared with the control group, in the model group, the error number in the Y-maze test, nerve function scores, apoptotic index, expression levels of Caspase-3 and Bax, and level of reactive oxygen species were significantly increased (P < 0.05), the expression of Bcl-2 and telomerase reverse transcriptase protein and mRNA, and level of superoxide dismutase were significantly decreased (P < 0.05). Compared with the model group, in the umbilical cord blood-derived mesenchymal stem cell 1 and 2 groups, the error number in the Y-maze test, nerve function score, apoptotic index, expression of Caspase-3 and Bax, and level of reactive oxygen species were significantly decreased (P < 0.05), while the expression of Bcl-2 and telomerase reverse transcriptase protein and mRNA and level of superoxide dismutase were significantly increased (P < 0.05). Compared with the umbilical cord blood-derived mesenchymal stem cell 2 group, in the umbilical cord blood-derived mesenchymal stem cell 1 group, the error number in the Y-maze test, nerve function score, apoptotic index, expression of Caspase-3 and Bax, and level of reactive oxygen species were significantly decreased (P < 0.05), while the expression of Bcl-2 and telomerase reverse transcriptase protein and mRNA and level of superoxide dismutase were significantly increased (P < 0.05). These findings indicate that umbilical cord blood-derived mesenchymal stem cell transplantation can effectively promote the neurological recovery in rats with cerebral infarction, and up-regulate the expression of telomerase reverse transcriptase in rat brain tissue. Moreover, earlier cell transplantation indicates better effects.

Key words: cerebral infarction, umbilical cord blood-derived mesenchymal stem cell, nerve function, telomerase reverse transcriptase, cell apoptosis

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