基础医学与临床 ›› 2012, Vol. 32 ›› Issue (7): 734-738.

• 研究论文 • 上一篇    下一篇

Genistein后处理介导eNOS激活及其对缺血后神经元的保护作用

涂静宜1,王瑞敏2,杨方3,朱莹1,周彩凤1,张茜1   

  1. 1. 河北联合大学医学实验研究中心
    2. 河北联合大学
    3. 华北煤炭医学院实验研究中心
  • 收稿日期:2011-06-21 修回日期:2011-11-21 出版日期:2012-07-05 发布日期:2012-06-20
  • 通讯作者: 王瑞敏 E-mail:ruimin-wang@163.com
  • 基金资助:
    NMDA受体诱导AKT/NOS/Nrf2/ARE信号通路-脑缺血后处理的重要保护机制

Genistein Postconditioning protects against I/R injury through eNOS activation in hippocampal CA1 region of rats

  • Received:2011-06-21 Revised:2011-11-21 Online:2012-07-05 Published:2012-06-20
  • Contact: Rui-min WANG E-mail:ruimin-wang@163.com

摘要: 目的 研究Genistein后处理 (Genistein Postconditioning, GPC) 对脑缺血再灌注后大鼠海马CA1区神经元的神经保护作用,及其对eNOS磷酸化水平的影响,从而揭示GPC神经保护作用可能的分子机制。方法 建立大鼠四动脉结扎全脑缺血模型,采用Western Blot技术检测大鼠海马CA1区eNOS、p-eNOS的表达;NeuN染色和TUNEL技术分别观察海马CA1区神经元的存活和凋亡样损伤。结果 1. Western Blot结果显示,与I/R组相比GPC后再灌注30min和3d p-eNOS水平显著升高,而eNOS蛋白表达在各个时间点没有明显变化。2. 激光扫描共聚焦显微镜技术结果显示,GPC组与对照组相比较,海马CA1区存活的神经元细胞明显增加,而凋亡样损伤的神经元显著减少;3. NOS抑制剂L-NAME不但有效降低p-eNOS水平,而且可显著减弱GPC诱导的神经保护作用。结论 Genistein后处理可抑制大鼠海马CA1区神经元凋亡,其机制可能与p-eNOS表达上调有关。

关键词: Genistein后处理, 脑缺血再灌注, p-eNOS, 海马CA1区

Abstract: Objective To investigate the neuroprotective role of genistein postconditioning (GPC) against cerebral ischemic injury and its effects on phosphorylation (activation) and protein expression of eNOS in hippocampal CA1 region of rats. Methods Cerebral ischemia was induced by four-vessel occlusion in rats and protein level of p-eNOS and eNOS were detected by Western Blotting. Additionally, NeuN staining was used to detect the survival neurons and apoptotic neurons of hippocampal CA1 region was observed using TUNEL analysis by Laser Scanning Confocal Microscope. Results In I/R groups, the number of NeuN-positive cell in hippocampal CA1 region was significant reduce accompanied with the increase of apoptotic neurons compared with sham groups. By contrast, treatment with Genistein after ischemia markedly protected the pyramidal neurons, as evidenced by the increase of NeuN-positive cell and the decrease of apoptotic neurons. Furthermore, L-NAME, an inhibitor of NOS abolished the neuroprotective role induced by genistein postconditioning. Additionally, immunoblot analysis showed that eNOS protein expression had no significant change in different times, while the level of p-eNOS significantly increased with two peaks occurring at 30min and 3d of reperfusion compared with I/R groups. L-NAME significantly suppressed enhance of p-eNOS induced by GPC at 3d of reperfusion. Conclusions GPC significantly prevents neuronal injury from global cerebral ischemia in the hippocampal CA1 region of rats. The possible mechanism is involved in the increase of p-eNOS expression.

Key words: Genistein Postconditioning, Cerebral ischemia/reperfusion, p-eNOS, Hippocampus CA1 region

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