文章摘要
幸享凤 ,王玉 ,张融融 ,秦新月 ,王恬竹.CRMP2真核表达质粒在大鼠缺血/再灌注脑皮质 转染效率的对比研究[J].神经损伤功能重建,2020,15(7):373-376
CRMP2真核表达质粒在大鼠缺血/再灌注脑皮质 转染效率的对比研究
Transfection Efficiency of CRMP2 Eukaryotic Expression Plasmid in Rat Cortex AfterIschemia/Reperfusion Injury
  
DOI:
中文关键词: Collapsin反应调节蛋白2  真核表达质粒  脑缺血再灌注  转染
英文关键词: collapsin response mediator protein 2  eukaryotic expression plasmid  cerebral ischemia/ reperfusion  transfection
基金项目:国家自然科学基金 (No. 81200900)
作者单位
幸享凤1 ,王玉2 ,张融融2 ,秦新月2 ,王恬竹2 1. 广州市花都区人 民医院神经内科 2. 重庆医科大学附 属第一医院神经内 科 
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中文摘要:
      目的:筛选Collapsin反应调节蛋白2(CRMP2)真核表达质粒转染大鼠脑皮质的适宜注射部位。方 法:成年雄性SD大鼠150只随机分为空白质粒(VP)组、侧脑室(LV)组、海马(H)组、皮质(C)组及嗅球 (OB)组。选定质粒注射部位后,20只大鼠随机分为假手术(sham)组、缺血再灌注(MCAO)组、缺血再灌 注+空白质粒(MCAO+GFP)组、缺血再灌注+CRMP2真核质粒干预(MCAO+CRMP2/GFP)组。大鼠脑缺 血/再灌注模型成功后,分别于相应4个部位立体定向注射CRMP2真核表达质粒/空白质粒,转染后24 h和 48 h,采用Western blot 及RT-PCR 检测缺血区皮质CRMP2蛋白及mRNA的表达;激光共聚焦检测GFP的 表达。TTC检测脑梗死体积,同时行神经功能评估。结果:不同部位注射CRMP2真核表达质粒后,皮质 CRMP2的表达均有增高,其中LV组及H组的转染效率明显高于C组及OB组(P<0.05),LV组与H组之间 差异无统计学意义(P>0.05)。与MCAO及(MCAO+GFP)组相比,CRMP2真核质粒干预之后能缩小脑梗 死体积,促进神经功能恢复(P<0.05)。结论:CRMP2真核表达质粒能够成功转染大鼠脑组织,使缺血区皮 质CRMP2蛋白及mRNA的表达明显增高。侧脑室的转染效率较其他三个部位更高。
英文摘要:
      To select the appropriate site of transfection of the collapsin response mediator protein 2 (CRMP2) eukaryotic expression plasmid in the cortex of rat brains. Methods: A total of 150 adult male Sprague-Dawley rats were randomly divided into groups vacant plasmid (VP), lateral ventricle (LV), hippocampus (H), cortex (C), and olfactory bulb (OB). After the appropriate site were chosen, 20 rats were divided into groups sham, middle cerebral artery occlusion (MCAO), (MCAO+GFP), (MCAO+CRMP2/GFP). The CRMP2 eukaryotic expression plasmid and vacant plasmid were respectively injected into the four different parts of MCAO/reperfusion model rats by stereotactic surgery; 24 h and 48 h after transfection, expression of CRMP2 protein and mRNA were tested by Western blot and RT-PCR assay and the expression of GFP was examined by laser scanning confocal microscope. TTC staining was employed to observe the cerebral infarct volume, and neurological function was also evaluated. Results: After injection of the plasmid, CRMP2 expression levels in the cortex increased; the expression levels in the LV and H groups were significantly higher than those in the C and OB groups (P<0.05) while there was no statistical difference between the LV and H groups (P>0.05). Compared to MCAO without plasmid treatment, CRMP2 eukaryotic plasmid intervention reduced cerebral infarction volume and promoted neural functional recovery (P<0.05). Conclusion: The CRMP2 eukaryotic expression plasmid can successfully transfect rat cortical tissue and lead to increased expression of CRMP2 proteins and mRNA in the ischemic cortical regions. The transfection rate is higher in the lateral ventricle than in the other three locations.
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