丁苯酞联合依达拉奉对大鼠局灶性脑缺血细胞中Drp1及Mfn2动态变化的影响及其保护机制  

Effect of butylphthalide combine edaravone on the dynamic change of Drp1 and Mfn2 in rats with focal cerebral ischemia and its protection mechanism

作　　者：陈婷[1] 李雪[1] 罗凡[1] 林占东 郭瑞芳[2] 

机构地区：[1]承德医学院研究生院,承德医学硕士研究生067000 [2]承德医学院附属医院老年病科,承德067000

出　　处：《医学研究生学报》2018年第3期284-289,共6页Journal of Medical Postgraduates

摘　　要：目的脑缺血后神经细胞发生氧化应激诱导细胞凋亡,从而打破线粒体的分裂融合的动态平衡。文章探讨丁苯酞联合依达拉奉对大鼠局灶性脑缺血细胞中Drp1及Mfn2动态变化的影响及其保护机制。方法将96只大鼠按随机数字表法分为缺血组、丁苯酞组、依达拉奉组、丁苯酞注射液联合依达拉奉组(联合用药组),每组又根据时间分为3、7、14 d的不同亚组,采用Longa-Zea线栓法建立大脑中动脉缺血模型。丁苯酞组、依达拉奉组及联合用药组分别从术前2 h及术后第1天开始腹腔注射依达拉奉10 m L/kg,丁苯酞注射液0.4 g/kg。脑缺血组在相同时间给予相同计量的等渗盐水。分别于第3、7、14天断头取左侧缺血区大脑皮质。采用神经功能学评分进行疗效评价。HE染色观察大脑皮质神经元形态结构,Western blot检测Drp1及Mfn2的蛋白含量,RT-PCR检测Drp1及Mfn2的mRNA的表达量。结果第3、7、14天,与脑缺血组比较,丁苯酞组、依达拉奉组、联合用药组评分降低(P<0.05);与联合用药组第3、7、14天神经功能缺损评分[(1.06±0.18)、(0.82±0.13)、(0.57±0.10)分]比较,丁苯酞组[(2.02±0.18)、(1.23±0.13)、(0.86±0.10)分]、依达拉奉组[(2.08±0.17)、(1.23±0.13)、(0.85±0.12)分]增加(P<0.05)。在相同时间水平下,与脑缺血组比较,丁苯酞组、依达拉奉组及联合用药组Drp1蛋白及mRNA表达量减少(P<0.05)。联合用药组Drp1蛋白及mRNA表达量减少(P<0.05)。与依达拉奉组比较,联合用药组Drp1蛋白及mRNA表达量减少(P<0.05);在相同时间水平下与脑缺血组比较,丁苯酞组、依达拉奉组及联合用药组Mfn2蛋白及mRNA表达量增多(P<0.05)。与丁苯酞组比较,联合用药组Mfn2蛋白及mRNA表达量增多(P<0.05)。与依达拉奉组比较,联合用药组Mfn2蛋白及mRNA表达量增多(P<0.05)。结论依达拉奉联合丁苯酞对脑缺血的保护作用效果强于单独使用,其机制可能与依达拉奉清除氧自Objective Apoptosis was induced by oxidative stress in nerve cells after cerebral ischemia.It further breaks the dynamic balance of mitochondrial division of fusion.This study aimed to investigate the effect of butylphthalide combined with edaravone treatment on the dynamic change of Drp1 and Mfn2 in rats with focal cerebral ischemia cells and its protection mechanism.Methods 96 rats were divided into 4 groups according to random number table.The 4 groups were ischemia group,butylphthalide group,edaravone group and butylphthalide combined with edaravone groups（combine group）,each group divided into three subgroups（3 d,7 d,14 d）.Longa-Zea suppository method is adopted to establish the middle cerebral artery occlusion（MCAO） model.Butylphthalide group,edaravone group and combine group were injected butylphthalide（0.4 g/kg） and/or edaravone（10 m L/kg） peritoneally 2 hours before surgery and 1 day after surgery.The same volume of isotonic saline was given at the same time of the other 3 groups in ischemia group.The cerebral cortex of the left ischemic region was obtained at the day 3,7,14.The evaluation of the curative effect was evaluated with neurological function score.HE staining was used to observe the cerebral cortex neuron morphological structure,protein and mRNA level of Drp1 and Mfn2 was measured by western blot and RT-PCR.Results At the day 3,7,and 14,the neurological function score was higher in ischemia group than the other 3 groups（P 0.05）.Compared with the combine group at day 3,7,and 14 [（1.06 ±0.18）,（0.82±0.13）,（0.57±0.10） ],the neurological function score was elevated in butylphthalide group [（2.02± 0.18）,（1.23±0.13）,（0.86±0.10） ] and edaravone group [（2.08±0.17）,（1.23±0.13）,（0.85±0.12） ]（P0.05）.At the same time point,compared with the ischemia group,Drp1 protein and mRNA levels were lower in the other 3 groups（P 0.05） while Mfn2 protein and mRNA levels were elevated（P0.05）.Compared with the butylphthalide group and edara

关 键 词：局灶性脑缺血 依达拉奉 丁苯酞注射液 线粒体 Drp1 MFN2 

分 类 号：R363.2[医药卫生—病理学]