螯合锌离子对局灶性脑缺血再灌注大鼠半暗带区低氧诱导因子-1α表达的影响

doi:10.3969/j.issn.1006-7795.2016.05.014

首都医科大学学报 ›› 2016, Vol. 37 ›› Issue (5): 626-630.doi: 10.3969/j.issn.1006-7795.2016.05.014

螯合锌离子对局灶性脑缺血再灌注大鼠半暗带区低氧诱导因子-1α表达的影响

房亚兰, 李森, 赵咏梅, 闫峰, 尹洁, 罗玉敏, 刘克建

首都医科大学宣武医院北京市老年病医疗研究中心神经变性病教育部重点实验室脑血管病转化医学北京市重点实验室, 北京 100053

收稿日期:2016-05-30 出版日期:2016-10-21 发布日期:2016-10-19
通讯作者: 赵咏梅 E-mail:yongmeizhao@hotmail.com
基金资助:
国家自然科学基金（81171242），北京市自然科学基金（7122036），脑血管病转化医学北京市重点实验室开放课题（2013NXGZ03）资助。

Effect of chelating zinc on expression of hypoxia inducible factor-1α in the penumbra of rats following focal cerebral ischemia/reperfusion

Fang Yalan, Li Sen, Zhao Yongmei, Yan Feng, Yin Jie, Luo Yumin, Liu Kejian

Xuanwu Hospital, Capital Medical University, Beijing Geriatric Medical Research Center, Key Laboratory of Neurodegenerative Diseases of Ministry of Education, Beijing Key Laboratory of Translational Medicine for Cerebrovascular Diseases, Beijing 100053, China

Received:2016-05-30 Online:2016-10-21 Published:2016-10-19
Supported by:
This study was supported by National Natural Science Foundation of China (81171242),Natural Science Foundation of Beijing (7122036),Open Project of Beijing Key Laboratory of Translational Medicine for Cerebrovascular Diseases (2013NXGZ03).

摘要/Abstract

摘要： 目的利用大脑中动脉梗塞（middle cerebral artery occlusion，MCAO）大鼠模型，观察脑缺血再灌注后脑组织低氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）表达的动态变化，并给予脑缺血大鼠锌离子螯合剂［N，N，N’，N ’-tetrakis（2-pyridylmethyl）ethylenediamine，TPEN］进行干预，研究锌离子对MCAO大鼠脑组织HIF-1α表达的影响，探讨锌离子介导脑缺血再灌注损伤的相关机制。方法采用数字表法将45只健康雄性Sprague-Dawley大鼠随机分为3组：假手术组（Sham）、MCAO组和MCAO+TPEN组（于MCAO前30 min腹腔注射TPEN，剂量为15 mg/kg），每组15只。使用线栓法制作大鼠右侧大脑中动脉缺血90 min再灌注模型。术中监测大鼠肛温及平均动脉压，使其维持在正常范围。分别于再灌注3、12和24 h处死大鼠，迅速取脑，用免疫组化染色检测大鼠脑组织冰冻切片内缺血半暗带区HIF-1α的表达水平，用免疫荧光双标对HIF-1α在缺血脑组织的表达进行细胞定位。结果假手术组大鼠未见HIF-1α染色阳性细胞，MCAO组大鼠脑组织缺血半暗带区HIF-1α的表达随再灌注时间延长逐渐增加（P<0.01）。给予15 mg/kg TPEN后，缺血再灌注各时间点大鼠脑组织缺血半暗带区内HIF-1α的表达比MCAO组明显减少（P<0.01）。缺血再灌注大鼠脑组织缺血半暗带区内，HIF-1α免疫荧光染色与神经元标志物NeuN免疫荧光染色共定位。结论大鼠局灶性脑缺血再灌注后，脑组织神经元内HIF-1α表达随再灌注时间延长递增。螯合细胞内锌离子下调缺血再灌注大鼠脑内HIF-1α浓度，提示锌离子可能通过促进HIF-1α表达介导脑缺血损伤。

关键词: 锌离子, 低氧诱导因子-1α, 脑缺血, 再灌注, 大鼠

Abstract: Objective To investigate the dynamic changes of hypoxia inducible factor-1α (HIF-1α) during reperfusion in a rat middle cerebral artery occlusion (MCAO)/reperfusion model and the changes of HIF-1α expression by removing zinc with zinc chelatorN,N,N',N'-tetrakis(2-pyridylmethyl) ethylenediamine, TPEN], and to explore the probable mechanism by which zinc involved in cerebral ischemia/reperfusion injury. Methods Forty-five male Sprague Dawley rats were divided into three groups randomly:Sham group, MCAO group, and MCAO+TPEN grouprats received TPEN (15mg/kg, i.p.) once 30 min before MCAO]. There were 15 rats in each group. MCAO operation was performed by using suture method. The rats underwent right MCAO for 90 min. The rectal temperature and mean arterial blood pressure were monitored to keep in normal range during the operation. At 3, 12 or 24 h after reperfusion, brains were removed. The expression and location of HIF-1α were detected by immunohistochemistry and immunofluorescent staining. Results 1) No HIF-1α positive cell was observed in the brain of Sham group rats. The number of HIF-1α positive cells in the penumbra of MCAO group showed time-dependent increasing tendency within 24 hours after reperfusion (P<0.01). 2) At 3, 12 and 24 h after reperfusion, the numbers of HIF-1α positive cells in the penumbra of MCAO+TPEN group rats decreased significantly compared with those of MCAO group rats (P<0.01). 3) HIF-1α-positive immunoreactive cells were colocalized with the general neuronal marker, NeuN, in MCAO rats. Conclusion The level of HIF-1α in neurons increased along with the extension of reperfusion time in focal cerebral ischemia/reperfusion rats. The expression of HIF-1α was decreased by removing intracellular zinc in MCAO rats, suggesting that zinc may be involved in the ischemia/reperfusion injury by promoting the expression of HIF-1α.

Key words: zinc, hypoxia inducible factor-1α (HIF-1α), brain ischemia, reperfusion, rats

中图分类号:

R743

房亚兰, 李森, 赵咏梅, 闫峰, 尹洁, 罗玉敏, 刘克建. 螯合锌离子对局灶性脑缺血再灌注大鼠半暗带区低氧诱导因子-1α表达的影响[J]. 首都医科大学学报, 2016, 37(5): 626-630.

Fang Yalan, Li Sen, Zhao Yongmei, Yan Feng, Yin Jie, Luo Yumin, Liu Kejian. Effect of chelating zinc on expression of hypoxia inducible factor-1α in the penumbra of rats following focal cerebral ischemia/reperfusion[J]. Journal of Capital Medical University, 2016, 37(5): 626-630.

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螯合锌离子对局灶性脑缺血再灌注大鼠半暗带区低氧诱导因子-1α表达的影响

Effect of chelating zinc on expression of hypoxia inducible factor-1α in the penumbra of rats following focal cerebral ischemia/reperfusion

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