天津医药 ›› 2018, Vol. 46 ›› Issue (2): 139-143.doi: 10.11958/20171240

• 实验研究 • 上一篇    下一篇

不同程度颅脑创伤大鼠外周血内皮祖细胞动态变化及伤后认知能力的研究

王计伟1, 2 , 王毅1 , 王东1 , 周源1 , 陈芳莲2 , 崔维韻2 , 刘丽2 , 张建宁1, 2△   

  1. 1天津医科大学总医院神经外科 (邮编300052); 2天津市神经病学研究所
  • 收稿日期:2017-11-09 修回日期:2017-12-14 出版日期:2018-02-15 发布日期:2018-02-15
  • 通讯作者: 张建宁 E-mail:jianningzhang@hotmail.com
  • 基金资助:
    天津市自然科学基金;国家自然科学基金资助面上项目;天津市应用基础与前沿技术研究计划;天津市高等学校基本科研业务费资助项目重点项目

Study on the dynamic changes of endothelial progenitor cells in peripheral blood and cognitive ability of rats with different degrees of traumatic brain injury

WANG Ji-wei 1, 2 , WANG Yi 1 , WANG Dong1 , ZHOU Yuan1 , CHEN Fang-lian2 , CUI Wei-yun2 , LIU Li 2 , ZHANG Jian-ning1, 2△   

  • Received:2017-11-09 Revised:2017-12-14 Published:2018-02-15 Online:2018-02-15
  • Contact: ZHANG JIanning E-mail:jianningzhang@hotmail.com

摘要: 摘要: 目的 观察不同程度颅脑创伤 (TBI) 大鼠外周循环血中内皮祖细胞 (EPCs) 和认知能力的动态变化。方法 采用随机数字表法将 28 只雄性 SD 大鼠分为假手术组和轻、 中、 重度 TBI 组, 每组 7 只。在前囟后 4 mm, 矢状缝右侧旁开 3 mm 处 (海马区域) 开圆形骨窗, 轻、 中、 重度 TBI 组分别采用 0.9、 2.1、 3.2 atm (1 atm=101.325 kPa) 的力度打击骨窗建立 TBI 模型, 分别在创伤前 (0 h) 及创伤后 3、 6、 24、 48、 72、 168、 240、 336 h 取内眦球后静脉丛血, 检测其循环血中 EPCs 水平, 并进行白细胞 (WBC) 和血小板 (PLT) 计数。TBI 后第 21~25 天, 采用 Morris 水迷宫定位巡航实验和空间探索实验比较各组大鼠的逃避潜伏期和目标象限百分率。结果 假手术组循环血中 EPCs 数量在整个实验过程中保持稳定。TBI 后 3 h, 轻、 中、 重度 TBI 组循环血中的 EPCs 数量 [(17.4±3.1) 个、(15.6±5.0) 个、(23.6± 3.0) 个] 低于假手术组 [(53.6±7.9) 个], 差异有统计学意义 (P<0.05); 在伤后 6 h, EPCs 数量迅速升高, 且轻度、 中度 TBI 组高于假手术组 (P<0.05), 约 48 h 后降至正常水平。整个实验过程中 EPCs 与 WBC、 PLT 的变化并不一致。定位巡航实验结果显示各组大鼠逃避潜伏期随着创伤时间的延长而缩短; 同期各 TBI 组逃避潜伏期均较假手术组延长。空间探索实验结果显示, 中、 重度 TBI 组大鼠停留在目标象限百分率明显低于假手术组和轻度 TBI 组。结论 颅脑创伤大鼠随着创伤程度加重, 其认知能力降低, 且循环血中 EPCs 的水平与病情严重程度有关, 可作为判断预后的标志物。

关键词: 关键词: 颅脑损伤, 内皮细胞, 认知障碍, 内皮祖细胞, 流式细胞术, 大鼠, Sprague-Dawley, 水迷宫实验

Abstract: Abstract: Objective To observe the dynamic changes of endothelial progenitor cells (EPCs) in peripheral blood and cognitive ability of rats with different degrees of traumatic brain injury (TBI). Methods A total of 28 male SD rats were randomly divided into 4 groups: sham group, mild traumatic brain injury group, moderate traumatic brain injury group and severe traumatic brain injury group, seven rats in each group. A hole was drilled on the right parietal skull (4.0 mm posterior from bregma and 3 mm lateral to the sagittal suture, hippocampal region) to expose the dura. Rats were subjected to different degrees of traumatic brain injury of 0.9, 2.1, 3.2 atm (1 atm=101.325 kPa). The dynamic changes of EPCs, white blood cell count (WBC) and platelets (PLT) in the circulating blood were measured before (0 h) and after TBI (3, 6, 24, 48, 72, 168, 240 and 336 h after trauma). Morris water maze (MWM) test was performed to record the escape latency and target quadrant change on day 21-25 after TBI in four groups. Results The number of circulating EPCs kept stable throughout the experiment in the sham group. The numbers of EPCs were significantly lower at 3 h after injury in mild, moderate and severe traumatic brain injury groups (17.4±3.1, 15.6±5.0 and 23.6±3.0) than those in the sham group (53.6±7.9, P<0.05). The numbers of EPCs at 6 h after injury were increased rapidly, and which were significantly higher in the mild and moderate TBI group than those in sham group (P<0.05). Then the number of EPCs dropped to the normal level on 48 h after injury. The changes of EPCs was inconsistent with the WBC and PLT during the whole experiment. The positioning cruise experiment showed that the escape latency shortened over time in each group. The escape latency was longer in TBI group than that in sham group during the same period. The spatial probe test showed that the percentages of the target quadrant were significantly lower in the moderate and severe TBI groups than those in the sham group and the mild TBI group. Conclusion With the severity of traumatic brain injury, the cognitive ability reduces in model rats. The level of endothelial progenitor cells in circulating blood is related to the severity of the traumatic brain injury, and can be used as a marker to judge the prognosis.

Key words: Key words: craniocerebral trauma, endothelial cells, cognition disorders, endothelial progenitor cells, flow cytometry, rats, Sprague-Dawley, water maze test