中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (9): 1585-1593.doi: 10.4103/1673-5374.237122

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

后肢远隔缺血后处理可保护卒中后血脑屏障的完整性

  

  • 收稿日期:2018-03-29 出版日期:2018-09-15 发布日期:2018-09-15
  • 基金资助:

    中国国家自然科学基金项目（30960107），四川省教育厅自然科学基金项目（14ZA0223）

Limb remote ischemic postconditioning protects integrity of the blood-brain barrier after stroke

Juan Li1, Xiao-Song Hu1, Fang-Fang Zhou1, Shuai Li1, You-Sheng Lin1, Wen-Qian Qi1, Cun-Fang Qi2, Xiao Zhang1   

  1. 1 Experiment Technology Center of Preclinical Medicine of Chengdu Medical College, Chengdu, Sichuan Province, China
    2 Department of Anatomy, Qinghai University, Xining, Qinghai Province, China
  • Received:2018-03-29 Online:2018-09-15 Published:2018-09-15
  • Contact: Cun-Fang Qi, Ph.D. or Xiao Zhang, Ph.D., 290683850@qq.com or Zhangxiao1985007@126.com
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 30960107; the Natural Science Foundation of the Education Department of Sichuan Province of China, No. 14ZA0223

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摘要：

血脑屏障结构的完整性对维持大脑内环境稳定至关重要，而脑梗死和脑水肿的发展与血脑屏障渗漏密切相关，因此，有研究认为保护血脑屏障可能是治疗急性脑卒中的一种有效的方法。为了验证这种可能性，实验以大脑中动脉闭塞再灌注建立卒中模型大鼠，于再灌注开始时即通过3个周期的双侧后肢10min缺血/10min再灌注诱导远隔缺血后处理干预治疗。以Zea longa法评估模型大鼠神经功能，以伊文思蓝渗漏评估血脑屏障通透性，以TTC染色评价脑梗死体积和脑水肿程度，以实时定量反转录PCR检测基质金属蛋白酶9和紧密连接蛋白5mRNA表达水平，以Western blot分析检测基质金属蛋白酶9和紧密连接蛋白5蛋白表达水平，以免疫组化分析基质金属蛋白酶9和紧密连接蛋白5免疫阳性细胞数量。24h后的结果显示，后肢远隔缺血后处理可减轻卒中大鼠血脑屏障破坏，缩小脑梗死体积和脑水肿体积，减少损伤区域基质金属蛋白酶9mRNA和蛋白表达以及阳性细胞数量，增加紧密连接蛋白5 mRNA和蛋白表达以及阳性细胞数量，显著改善其神经功能。验证后肢远隔缺血后处理可通过抑制急性脑缺血再灌注诱导的基质金属蛋白酶9和紧密连接蛋白5的表达变化，减轻血脑屏障损伤，从而减轻缺血性损伤，实现对大脑的保护作用。

orcid:0000-0002-3311-8166(Cun-Fang Qi)

关键词: 后肢远隔缺血后处理, 大脑中动脉闭塞, 脑缺血再灌注, 血脑屏障, 急性脑缺血, 卒中, 基质金属蛋白酶9, 紧密连接蛋白5

Abstract:

Integrity of the blood-brain barrier structure is essential for maintaining the internal environment of the brain. Development of cerebral infarction and brain edema is strongly associated with blood-brain barrier leakage. Therefore, studies have suggested that protecting the blood-brain barrier may be an effective method for treating acute stroke. To examine this possibility, stroke model rats were established by middle cerebral artery occlusion and reperfusion. Remote ischemic postconditioning was immediately induced by three cycles of 10-minute ischemia/10-minute reperfusion of bilateral hind limbs at the beginning of middle cerebral artery occlusion reperfusion. Neurological function of rat models was evaluated using Zea Longa’s method. Permeability of the blood-brain barrier was assessed by Evans blue leakage. Infarct volume and brain edema were evaluated using 2,3,5-triphenyltetrazolium chloride staining. Expression of matrix metalloproteinase-9 and claudin-5 mRNA was determined by real-time quantitative reverse transcription-polymerase chain reaction. Expression of matrix metalloproteinase-9 and claudin-5 protein was measured by western blot assay. The number of matrix metalloproteinase-9- and claudin-5-positive cells was analyzed using immunohistochemistry. Our results showed that remote ischemic postconditioning alleviated disruption of the blood-brain barrier, reduced infarct volume and edema, decreased expression of matrix metalloproteinase-9 mRNA and protein and the number of positive cells, increased expression of claudin-5 mRNA and protein and the number of positive cells, and remarkably improved neurological function. These findings confirm that by suppressing expression of matrix metalloproteinase-9 and claudin-5 induced by acute ischemia/reperfusion, remote ischemic postconditioning reduces blood-brain barrier injury, mitigates ischemic injury, and exerts protective effects on the brain.

Key words: nerve regeneration, remote ischemic postconditioning, middle cerebral artery occlusion, cerebral ischemia/reperfusion, bloodbrain barrier, acute cerebral ischemia, stroke, matrix metalloproteinase-9, claudin-5, neural regeneration