中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (11): 1773-1778.doi: 10.4103/1673-5374.194747

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

未参与局灶性脑缺血后再灌注早期活性氧生成的NADPH氧化酶2

  

  • 收稿日期:2016-09-20 出版日期:2016-11-30 发布日期:2016-11-30
  • 基金资助:

    深圳市科技计划项目（JCYJ20150330102401097, KQCX20140521101427034, JCYJ20140414170821291）；中国博士后科学基金（2015M572388）

NADPH oxidase 2 does not contribute to early reperfusion-associated reactive oxygen species generation following transient focal cerebral ischemia

Yuan Zhang1, 2, 3, Ting Wang1, 2, 4, Ke Yang1, 2, 4, Ji Xu1, 2, Jian-ming Wu5, Wen-lan Liu1, 2, *   

  1. 1 Central Laboratory, Shenzhen Second People’s Hospital, First Afliated Hospital of Shenzhen University, Shenzhen, Guangdong Province, China 2 Shenzhen Key Laboratory of Neurosurgery, Shenzhen Second People’s Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong Province, China 3 Department of Pathophysiology, Baotou Medical College, Baotou, Inner Mongolia Autonomous Region, China 4 Graduate School of Guangzhou Medical University, Guangzhou, Guangdong Province, China 5 Department of Neurosurgery, Shenzhen Second People’s Hospital, First Afliated Hospital of Shenzhen University, Shenzhen, Guangdong Province, China
  • Received:2016-09-20 Online:2016-11-30 Published:2016-11-30
  • Contact: Wen-lan Liu, M.D., Ph.D., wlliu@szu.edu.cn.
  • Supported by:

    The work was financially supported by grants from Shenzhen Science and Technology Innovation Commission of China, No. JCYJ20150330102401097, KQCX20140521101427034, JCYJ20140414170821291, and China Postdoctoral Science Foundation, No. 2015M572388.

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摘要：

活性氧的过度产生可明显加重缺血再灌注损伤以及血流恢复后脑组织的病理改变，但活性氧产生的酶促反应机制目前仍有待明晰。我们设计了观察含NADPH氧化酶的Nox2在缺血再灌后脑组织中的表达和活性的实验，以明确其机制。将雄性SD大鼠的大脑中动脉闭塞90 min后，随即再灌注3 h或22.5 h，利用荧光定量PCR和蛋白质印迹方法分别检测缺血脑组织中Nox2的mRNA和蛋白表达水平，采用光泽精荧光测定Nox的酶活性。结果显示，再灌注22.5 h的脑组织中Nox2 mRNA和蛋白的表达量分别增加了3.7倍和3.6倍，再灌注3 h后脑组织中Nox2 mRNA和蛋白的表达并没有发生明显改变，同时，在2个再灌注时间点的缺血脑组织中，NADPH氧化酶发生了类似改变。实验结果表明。Nox2可能与早期再灌注相关的活性氧产生无关，而是延时再灌注期间活性氧产生的重要来源。 

orcid: 0000-0003-2130-4065 (Wen-lan Liu)

关键词: 神经再生, 脑损伤, NADPH氧化酶, 脑缺血, 非吞噬细胞氧化酶家族, 活性氧, 再灌注, 时间依赖性, 脑卒中, 血流, 大鼠, 早期阶段

Abstract:

Excess production of reactive oxygen species (ROS) critically contributes to occurrence of reperfusion injury, the paradoxical response of ischemic brain tissue to restoration of cerebral blood ?ow. However, the enzymatic sources of ROS generation remain to be unclear. Tis study examined Nox2-containing NADPH oxidase (Nox2) expression and its activity in ischemic brain tissue following post-ischemic reperfusion to clarify the mechanism of enzymatic reaction of ROS. Male Sprague-Dawley rats were subjected to 90-minute middle cerebral artery occlusion, followed by 3 or 22.5 hours of reperfusion. Quantitative reverse transcriptase PCR and western blot assay were performed to measure mRNA and protein expression of Nox2. Lucigenin ?uorescence assays were performed to assess Nox activity. Our data showed that Nox2 mRNA and protein expression levels were signifcantly increased (3.7-fold for mRNA and 3.6-fold for protein) in ischemic brain tissue at 22.5 hours but not at 3 hours following post-ischemic reperfusion. Similar results were obtained for the changes of NADPH oxidase activity in ischemic cerebral tissue at the two reperfusion time points. Our results suggest that Nox2 may not contribute to the early burst of reperfusion-related ROS generation, but is rather an important source of ROS generation during prolonged reperfusion.

Key words: nerve regeneration, NADPH oxidase, cerebral ischemia, Nox family, reactive oxygen species, reperfusion, central nervous system, stroke, blood ?ow, neural regeneration