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黄芪注射液对脑缺血再灌注大鼠海马神经元形态学和Apaf-1表达的影响

刘瑞1,高维娟2,钱涛1,王利1   

  1. 1. 承德医学院病理生理学教研室
    2. 河北化工医药职业技术学院
  • 收稿日期:2013-03-21 修回日期:2013-08-27 出版日期:2013-12-15 发布日期:2013-12-15
  • 通讯作者: 高维娟

The Effects of Astragalus Injection on the Morphology and Expression of Apaf-1 in Hippocampal Neurons after Cerebral Ischemia Reperfusion in Rats

LIU Rui 1,GAO Wei juan2,QIAN Tao 1,WANG Li 1   

  1. 1. Department of Pathophysioloyg, Chengde Medical College
    2. Hebei Chemical and Pharmaceutical College
  • Received:2013-03-21 Revised:2013-08-27 Published:2013-12-15 Online:2013-12-15
  • Contact: GAO Wei juan

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摘要：

【摘要】  目的  观察黄芪注射液对脑缺血再灌注大鼠海马神经元形态学和Apaf-1表达的影响。  方法  SD大鼠随机分为假手术组、脑缺血再灌注组（再灌注组）和黄芪注射液处理组（实验组）。采用Pulsinelli四血管阻断法建立大鼠脑缺血再灌注模型，实验组于术前30min腹腔注射黄芪注射液6mL/kg，每24h注射1次，各组于再灌注后24h断头取脑。采用HE染色，光镜下观察海马组织病理学改变；透射电镜下观察海马神经元超微结构变化；免疫组织化学法和Western blot法检测大鼠海马组织凋亡蛋白酶激活因子-1（Apaf-1）蛋白表达的变化。  结果  与假手术组比较，再灌注组大鼠海马神经元出现胞核及线粒体损伤，并且Apaf-1蛋白的表达明显升高（免疫组化：0.024±0.001vs0.109±0.011；Western blot法：0.270±0.018vs0.894±0.072，均P＜0.01），与再灌注组比较，实验组大鼠海马神经元胞核及线粒体损伤明显减轻，而Apaf-1蛋白的表达明显降低（免疫组化：0.048±0.005；Western blot法0.392±0.046，均P＜0.01）。   结论  黄芪注射液可减轻脑缺血再灌注大鼠海马神经元的损伤，其作用机制可能与抑制Apaf-1蛋白表达有关

关键词: 再灌注损伤, 黄芪, 海马, 细胞凋亡, 细胞凋亡蛋白酶激活因子1

Abstract:

[Abstract]   Objective   To investigate the effects of astragalus injection on the morphology and expression of Apaf-1
in hippocampal neurons after cerebral ischemia reperfusion in rats.   Methods   The male SD rats were randomly divided into 3groups, sham-operated group, cerebral ischemia-reperfusion group (reperfusion group) and astragalus injection intervention group (experiment group). The global cerebral ischemia-reperfusion rat model was established by Pulsinelli four-vessel occlusion method. The astragalus injection group was intraperitoneally injected with astragalus6mL/kg,30mins before surgery and repeated every24h. Rat brains were removed24h after reperfusion in each group. HE staining was used to observe the pathological changes of the hippocampal neurons under the light microscope. The ultrastructural changes of hippocam?pal neurons were observed by transmission electron microscopy. Immunohistochemistry and Western blot methods were used to measure the expression of apoptotic protease activating factor-1(Apaf-1) protein.   Results   Compared with sham-operated group, nuclear and mitochondrial damage was found in reperfusion group, and the expression of Apaf-1protein increased obviously in hippocampus（Immunohistochemistry result:0.024±0.001vs0.109±0.011;Western blot result: 0.270±0.018vs0.894±0.072, P＜0.01). Compared with reperfusion group, the damage in nuclear and mitochondria was relieved obviously in experiment group, and the expression of Apaf-1protein in hippocampus was significantly decreased (Immunohistochemistry result:0.048±0.005;Western blot result: 0.392±0.046, P＜0.01).   Conclusion   Astragalus injection can reduce pathological damage of hippocampal neurons after cerebral ischemia and reperfusion in rats, and the mechanism is related with inhibiting of Apaf-1protein.

Key words: reperfusion injury, astragalus membranaceus, hippocampus, apoptosis, Apaf-1