《中国康复理论与实践》

《中国康复理论与实践》

• 基础研究 • 上一篇    下一篇

高压氧对脑外伤外源性骨髓间充质干细胞归巢的影响①

刘杨1，丁政1，唐朝正1，周苏键2，卢晓欣2，彭慧平1   

  1. 1. 福建中医药大学，福建福州市350100；2. 南京军区福州总医院康复医学科，福建福州市350025。
  • 出版日期:2016-07-25 发布日期:2016-09-22

Effects of Hyperbaric Oxygen on Homing of Exogenous Bone Marrow Mesenchymal Stem Cells after Traumatic Brain Injury

LIU Yang1, DING Zheng1, TANG Chao-zheng1, ZHOU Su-jian2, LU Xiao-xin2, PENG Hui-ping1   

  1. 1. Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350100, China; 2. Department of Rehabilitation Medicine, Fuzhou General Hospital of Nanjing Command, PLA, Fuzhou, Fujian 350025, China
  • Published:2016-07-25 Online:2016-09-22

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摘要： 目的探讨高压氧治疗对脑外伤大鼠外源性骨髓间充质干细胞(BMSCs)归巢的影响。方法采用Ficoll 密度梯度离心法分离、培养BMSCs，应用流式细胞仪对第三代BMSCs表面标志CD29、CD90、CD45、CD11b进行鉴定，细胞膜荧光探针CM-DiI 对BMSCs进行示踪处理。36 只Sprague-Dawley 大鼠分为假手术组(A 组，n=6)、模型组(B 组，n=6)、BMSCs移植组(C 组，n=12)、高压氧+BMSCs组(D组，n=12)，分别于移植后1 d、3 d 取脑组织标本，冰冻切片，荧光显微镜下观察示踪后BMSCs归巢的数量和部位；Western blotting 检测基质细胞衍生因子1(SDF-1)及其受体CXCR4 的蛋白表达。结果受伤侧大脑半球，尤其是受损脑组织周围，荧光标记BMSCs更为集中；同一时间点D组BMSCs归巢数量明显高于C组(P<0.01)；C、D组3 d 时BMSCs归巢数量均明显高于1 d 时(P<0.01)。D组SDF-1、CXCR4 的蛋白表达水平高于C组(P<0.05)。结论高压氧治疗可以促进外源性BMSCs归巢至大鼠受损脑组织，其作用与SDF-1/CXCR4 信号轴的表达增强相关。

关键词: 脑外伤, 高压氧, 骨髓间充质干细胞, 归巢, 大鼠

Abstract: Objective To investigate the effects of hyperbaric oxygen (HBO) on homing of exogenous bone marrow mesenchymal stem cells (BMSCs) after traumatic brain injury (TBI) in rats. Methods BMSCs were isolated and cultured with Ficoll density gradient centrifugation, and the surface markers (CD29, CD90, CD45, CD11b) of the third generation were identified with flow cytometry. The authenticated BMSCs were processed by the cell membrane fluorescent probe CM-DiI before transplantation. Thirty-six Sprague-Dawley rats were divided into Sham group (n=6), TBI group (n=6), BMSCs group (n=12), HBO+BMSCs group (n=12). The number and locations of homing of tracing BMSCs were observed under fluorescent microscope after frozen sections, and the expression of stromal cell-derived factor 1 (SDF-1) and CXC chemokine receptor type 4 (CXCR4) proteins were detected with Western blotting one and three days after BMSCs transplantation. Results The fluorescence-labeled BMSCs focused on the injured hemisphere, especially around the damaged brain tissue. The number of homing was more in HBO+BMSCs group than in BMSCs group at the same time (P<0.01), and increased in both groups three days after transplantation compared with those of one day after transplantation (P<0.01). The expression of SDF-1 and CXCR4 protein were more in HBO+ BMSCs group than in BMSCs group (P<0.05). Conclusion HBO can promote the exogenous BMSCs homing to damaged brain tissue in rats after traumatic brain injury, which is related to the enhancement of SDF-1/CXCR4 axis.

Key words: traumatic brain injury, hyperbaric oxygen, bone marrow mesenchymal stem cells, homing, rats