miR-335在局灶性脑缺血大鼠脑组织中的表达变化及作用机制研究  

Expression changes of miR-335 in focal cerebral ischemia tissues in rats and their related mechanisms

作　　者：李晓云[1] 褚静[1] 牛建一[1] 陈红兵[1] 王立新[1] 

机构地区：[1]潍坊市益都中心医院神经内科,262500

出　　处：《中华神经医学杂志》2017年第6期570-575,共6页Chinese Journal of Neuromedicine

摘　　要：目的探讨miR-335在局灶性脑缺血大鼠脑组织中的表达变化及作用机制。方法50只成年健康雄性SD大鼠按随机数字表法分为假手术组、模型组、miR-335转染组、阴性对照组和空白质粒组，每组10只。后4组采用Longa线栓法制备成永久性局灶性脑缺血模型。模型制备成功后假手术组和模型组将等体积生理盐水注入侧脑室，miR-335转染组将重组质粒pcDNA3．1-miR-335和硬脂酰胺（SA）脂质体混合物注入侧脑室，阴性对照组将重组质粒pcDNA3．1．阴性对照和SA脂质体混合物注入侧脑室，空白质粒组将pcDNA3.1（＋）质粒和SA脂质体混合物注入侧脑室。于脑缺血后24h时，采用Zea-Longa评分方法对各组大鼠进行神经功能评分，采用TTC染色测量各组大鼠缺血脑组织体积，采用实时荧光定量PCR（RTFQPCR）技术检测各组大鼠缺血脑组织中miR-335的表达，采用Westernblottign检测各组大鼠缺血脑组织中血小板内皮细胞粘附分子1（CD31）和血管内皮生长因子（VEGF）蛋白的表达。结果与假手术组相比。模型组、miR-335转染组、阴性对照组和空白质粒组大鼠神经功能评分和缺血脑组织体积均明显增加．差异均有统计学意义（P〈0．05）；与模型组相比，miR-335转染组大鼠神经功能评分和缺血脑组织体积均明显降低．差异均有统计学意义（P〈0．05）。与假手术组相比，模型组、miR-335转染组、阴性对照组和空白质粒组大鼠缺血脑组织中miR-335相对表达量均明显降低，差异均有统计学意义（P〈0．05）；与模型组相比，miR-335转染组大鼠缺血脑组织中miR-335相对表达量明显升高．差异有统计学意义（P〈0．05）。与假手术组相比，模型组、miR-335转染组、阴性对照组和空白质粒组大鼠缺血脑组织中CD31和VEGF蛋白相对表达量均明显降低，差异均有统计学意义（P〈0．05）；与模型组相比，miR一-335转染组大Objective To investigate the expression changes of miR-335 in rats with focal cerebral ischemia and their mechanisms. Methods Fifty adult healthy male SD rats were randomly divided into sham-operated group, model group, miR-335 transfection group, negative control group and blank plasmid group （n=10）. The focal cerebral ischemia rat models in the later 4 groups were constructed by Longa method. After model making, rats in the miR-335 transfection group were injected recombinant plasmid pcDNA3A-miR-335 and stearamide （SA） liposome mixture into the lateral ventricle, rats in the negative control group were injected pcDNA3.1-negative control and SA liposome mixture into the lateral ventricle, rats in the blank plasmid group were injected pcDNA3.1 （＋） and SA liposome mixture into the lateral ventricle, and rats in the sham-operated group and model group were injected the same volume of saline. Twenty-four h after ischemia, the neurological deficit of these rats were assessed by Zea-Longa scale. These rats were sacrificed, and the brain infarct volumes were measured by TTCstaining; the miR-335 expression in cerebral ischemia tissues of rats was detected by using real-time fluorescence quantification PCR （RTFQ PCR）; the protein expressions of cell adhesion molecule 1 （CD31） and vascular endothelial growth factor （VEGF） in cerebral ischemia tissues of rats were detected by Western blotting. Results As compared with those in the sham-operated group, the neurological function scale scores and brain infarct volumes of rats in model group, miR-335 transfection group, negative control group and blank plasmid group were significantly increased （P〈0.05）; the neurological function scale scores and brain infarct volumes of rats in miR-335 transfection group were significantly decreased as compared with those in the model group （P〈0.05）. As compared with those in the sham-operated group, the relative miR-335 expression level in the cerebral ischemia tissues of the model group, miR-335 transfe

关 键 词：局灶性脑缺血 miR-335 血小板内皮细胞粘附分子1 血管内皮生长因子 

分 类 号：R743.3[医药卫生—神经病学与精神病学]