中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (40): 7493-7496.doi: 10.3969/j.issn.1673-8225.2010.40.018

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

NMDA受体拮抗剂剂量与脑缺血再灌注大鼠海马内源性神经干细胞的增殖

任铭新1,2，周  立2，李娜娜2，冯志博2，臧卫东1，袁国艳3   

  1. 1郑州大学基础医学院人体解剖学教研室，河南省郑州市  450000； 2新乡医学院人体解剖学教研室，河南省新乡市   453003；3新乡医学院第一附属医院神经外科，河南省卫辉市    453100
  • 出版日期:2010-10-01 发布日期:2010-10-01
  • 通讯作者: 袁国艳，硕士，主治医师，新乡医学院第一附属医院神经外科，河南省卫辉市453100 hnlyrmx@163.com
  • 作者简介:任铭新★，男，1979年生，河南省周口市人，汉族，郑州大学在读硕士。 hnlyrmx@163.com
  • 基金资助:

    2009年度新乡医学院第七批省级重点学科开放课题资助项目(ZD200923)。

N-methyl-D-aspartic acid receptor antagonist dose and proliferation of endogenous neural stem cells in the hippocampus of rats with cerebral ischemia/reperfusion

Ren Ming-xin 1,2, Zhou Li2, Li Na-na2, Feng Zhi-bo2, Zang Wei-dong1, Yuan Guo-yan3   

  1. 1 Department of Human Anatomy, Basic Medical College, Zhengzhou University, Zhengzhou  450000, Henan Province, China; 2 Department of Human Anatomy, Xinxiang Medical University, Xinxiang  453003, Henan Province, China; 3 Department of Neurosurgery, First Affiliated Hospital, Xinxiang Medical University, Weihui  453100, Henan Province, China
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Yuan Guo-yan, Master, Attending physician, Yuan Guo-yan, Master, Attending physician, Department of Neurosurgery, First Affiliated Hospital, Xinxiang Medical University, Weihui 453100, Henan Province, China hnlyrmx@163.com
  • About author:Ren Ming-xin★, Studying for master’s degree, Department of Human Anatomy, Basic Medical College, Zhengzhou University, Zhengzhou 450000, Henan Province, China; Department of Human Anatomy, Xinxiang Medical University, Xinxiang 453003, Henan Province, China hnlyrmx@163.com
  • Supported by:

    the Seventh Batch Provincial-level Key Subject Opening Topic of Xinxiang Medical University in 2009, No. ZD200923*

PDF

326

被引次数

4

摘要：

背景：脑缺血再灌注后，过度释放的兴奋性氨基酸可通过NMDA受体激活内源性神经干细胞，促使其增殖、分化，修复神经细胞，但同时也导致细胞内钙离子超载，引起神经细胞的损伤。通过控制NMDA受体活化的程度，刺激内源性神经干细胞激活的同时把损伤减到最小。
目的：观察NMDA受体拮抗剂MK-801质量浓度对脑缺血再灌注大鼠海马内源性神经干细胞增殖的影响。
方法：健康雄性SD大鼠54只随机数字表法分为对照组(不进行任何处理)、手术组(缺血模型制作)及不同剂量MK-801组。采用大鼠四条血管阻断方法(Pulsinelli-4VO法)制备大鼠全脑缺血再灌注模型。不同浓度MK-801组在模型制作前30 min按照0.2，0.4，0.6，0.8，1.0 ，1.2 mg/kg侧脑室注射MK-801。Western-blot、RT-PCR技术检测各组nestin蛋白及mRNA水平及表达。
结果与结论：MK-801剂量在0.8 mg/kg以下时，nestin mRNA及蛋白的表达差异无显著性意义(P > 0.05)，呈现高表达；当剂量为0.8 mg/kg时，可明显抑制内源性神经干细胞增殖；在0.8mg/kg以上，对神经干细胞的抑制作用随着剂量的升高呈递增趋势；在1.2 mg/kg时，大鼠有躁动、共济失调等严重不良反应。nestin mRNA表达结果与蛋白表达趋势相吻合。说明MK-801的剂量与脑梗死后神经功能的修复有一定的关系，实验中0.6 mg/kg是一个比较合适的实验应用剂量，在此剂量下既可使MK-801减少兴奋性氨基酸对神经元的毒性作用，保护神经细胞，又使内源性神经干细胞的增殖不受较大影响。

关键词: 脑缺血, 神经干细胞, NMDA受体拮抗剂, 海马, 增殖

Abstract:

BACKGROUND: After cerebral ischemia-reperfusion, excessive excitatory amino acids activate N-methyl-D-aspartic acid (NMDA) receptor which repairs nerve cells by the proliferation and differentiation of endogenous neural stem cells (NSCs), but damages nerve cells by intracellular calcium overload at the same time. Controlling NMDA receptor activation may activate endogenous NSCs and minimize damage simultaneously.
OBJECTIVE: To investigate the effect of NMDA receptor antagonist MK-801 on endogenous neural stem cell proliferation of reperfused rat hippocampus.
METHODS: A total of 54 healthy male Sprague-Dawley rats were randomized into control group (without any treatment), operation group (establish ischemia model), various doses of MK-801 group. Four-vessel occlusion (Pulsinelli-4VO method) was used to establish global cerebral ischemia/reperfusion model. Different doses of MK-801 were injected into lateral ventricle 30 minutes before cerebral ischemia according to 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 mg/kg. Western-blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect nestin protein and mRNA level expression in each group.
RESULTS AND CONCLUDION: When MK-801 was under 0.8 mg/kg, no significant differences in the protein and mRNA expression of nestin were detected among groups (P > 0.05), with the presence of high expression. When MK-801 was equal to 0.8 mg/kg, MK-801 significantly suppressed proliferation of endogenous NSCs. When MK-801 was more than 0.8 mg/kg, inhibitory effect was increased with increased concentration. At 1.2 mg/kg, rats developed adverse reaction such as restlessness and ataxia. The nestin mRNA expression results were consistent with the tendency of protein expression, which suggested that MK-801 dose is associated with the repair of nerve function following cerebral infarction. 0.6 mg/kg is a suitable dose during this test. This concentration can make MK-801 reduce the toxic effect of excitatory amino acids on neurons, can protect neural cells, and cannot greatly affect proliferation of endogenous NSCs.

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