基础医学与临床 ›› 2017, Vol. 37 ›› Issue (6): 802-807.

• 研究论文 • 上一篇    下一篇

人脂肪间充质干细胞来源的外排体促进大鼠创伤性脑损伤后神经功能恢复

陈云飞1,马百涛2,薛春玲1,韩钦3,魏俊吉4,赵春华1,常健博5   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院 北京协和医院
    3. 中国医学科学院基础医学研究所组织工程中心
    4. 中国医学科学院北京协和医院神经外科
    5. 中国医学科学院北京协和医学院北京协和医院
  • 收稿日期:2017-03-23 修回日期:2017-04-24 出版日期:2017-06-05 发布日期:2017-05-26
  • 通讯作者: 韩钦 E-mail:hanqinhanqin@126.com
  • 基金资助:
    间充质干细胞通过分泌的Exosome 促进脑缺血大鼠神经修 复的机制的研究

Exosome extracted from hAMSCs promotes neurological function recovery after taumatic brain injury in rats

  • Received:2017-03-23 Revised:2017-04-24 Online:2017-06-05 Published:2017-05-26
  • Contact: Qin HAN, E-mail:hanqinhanqin@126.com

摘要: 目的:研究人脂肪间充质干细胞(AMSCs)来源的外排体对创伤性脑损伤(TBI)的治疗作用及其可能的机制。方法:分离健康成人脂肪MSCs,通过超滤法提取外排体。将大鼠分成:假手术组,PBS对照组,MSC治疗组,exosomes治疗组。于TBI建模 24h后,治疗组分别沿损伤边缘区局部注射,PBS 30μL, MSC 2ⅹ105个cells/只,exosomes 25μg总蛋白量/只,总体积30μL。在建模前和TBI后1、3、7、10、13、16、和30d测试所有大鼠的mNSS评分和前肢踩空试验。3和7d处死大鼠,提取大鼠脑组织总RNA,实时定量PCR检测大鼠炎性因子TNF-α和IL-1β的表达,30d处死大鼠,tunel-neun双标免疫荧光检测TBI后神经元凋亡。结果:外排体的治疗显著促进神经功能的恢复,治疗效果与MSC治疗效果相当,其机制可能是通过抑制大鼠TBI后急性炎性反应,减少神经元凋亡。

关键词: 间充质干细胞, 外泌体, 创伤性脑损伤, 细胞治疗, 动物模型

Abstract: Objective: To investigate the therapeutic effect of exosomes extracted from human adipose-derived mesenchymal stem cells on traumatic brain injury (TBI) and its possible mechanism. Methods: Mesenchymal stem cells(MSCs) were isolated from healthy human adipose tissue and the exosomeswere extracted by ultrafiltration. Rats were divided into four groups: sham group, PBS control group, MSCs treatment group and exosomes treatment group. 24 h After TBI, the treatment group was injected locally along the lesion area, 30μL of PBS, 2 ⅹ105MSC, 25μg protein of exosomes respectively, the total volume was30μL. We performed the Modified Neurological Severity Score(mNSS) and the forelimb Foot-Fault Test in all rats beforeinjury and at 1, 3, 7, 10, 13, 16, 21, and 30 days after TBI. The rats were sacrificed at 3 and 7 days after TBI respectively,total RNA was extracted from rat brain tissue. The expression of TNF-α and IL-1β were detected by quantitative PCR. The rats were also killed at 30 days after TBI for testing the neuronal apoptosis in lesion area by tunel-neun double immunofluorescence. Results:exosomes treatment significantly promoted the recovery of neurological deficits caused by TBI,and the therapeutic effect was similar to MSCs, its possible mechanism may be the inhibition of the acute inflammation and the reducing of the neurons apoptosis after TBI.

Key words: mesenchymal stem cells (MSCs), Exosome, traumatic brain injury (TBI), cell therapy, animal models