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络瘀通胶囊对大鼠脑缺血-再灌注损伤的作用
引用本文:王丽晔,;赵海苹,;王荣亮,;张陈诚,;刘小蒙,;王宁群,;罗玉敏,;陈志刚.络瘀通胶囊对大鼠脑缺血-再灌注损伤的作用[J].中国脑血管病杂志,2014(12):650-655.
作者姓名:王丽晔  ;赵海苹  ;王荣亮  ;张陈诚  ;刘小蒙  ;王宁群  ;罗玉敏  ;陈志刚
作者单位:[1]北京中医药大学东方医院脑一科,100078; [2]首都医科大学宣武医院脑血管病研究室北京市老年病医疗研究中心,100078;
基金项目:北京市中医药科技项目(JJ2012-07);首都医科大学中医药研究专项(13ZY10)
摘    要:目的观察并探讨络瘀通胶囊对大鼠脑缺血-再灌注后神经功能的影响及其保护机制。方法采用改良Longa法制备大鼠大脑中动脉阻塞(MCAO)模型,缺血1.5 h后进行再灌注。选取10只大鼠作为假手术组,将40只造模成功的雄性SD大鼠按照随机数字表法分为4组,每组10只:模型组、络瘀通中剂量组(LYTM组)、络瘀通高剂量组(LYTH组)及胞磷胆碱钠组(CS组)。分别于造模后3 d及7 d采用12分评分及前肢踩空实验评价大鼠神经功能,应用蛋白免疫印迹法(Western blot)检测造模3 d后大鼠缺血侧及假手术组同侧脑组织中脑源性神经营养因子(BDNF)、碱性成纤维生长因子(b-FGF)及磷酸化蛋白激酶/蛋白激酶(p-AKT/AKT)的表达,及造模后7 d同侧脑组织中Caspase-12的表达并进行组间比较。结果 (1)神经功能评分:造模后3 d各组12分评分及前肢踩空实验评分差异均无统计学意义(均P0.05);造模后7 d,LYTM组、LYTH组、CS组较模型组均明显改善(均P0.05)。(2)Western blot结果显示:1模型组BDNF及b-FGF表达明显少于假手术组(均P0.05);与模型组比较,LYTM组、LYTH组、CS组均可上调BDNF和b-FGF表达,以LYTH组作用最明显(P0.01)。2与假手术组比较,模型组p-AKT/AKT表达明显减少(P0.05);与模型组比较,LYTM组、LYTH组、CS组p-AKT/AKT表达增加,以LYTH组和CS组明显,差异均有统计学意义(均P0.05)。3在Caspase-12的表达中,模型组裂解后Caspase-12表达较假手术组明显升高,差异有统计学意义(P0.05)。与模型组比较,LYTM组、LYTH组Caspase-12前体(pro Caspase-12)和裂解后Caspase-12表达有减少趋势,但差异无统计学意义(均P0.05);CS组Caspase-12前体和裂解后Caspase-12表达水平明显低于模型组(P0.05)。结论络瘀通胶囊可通过促进神经元存活及再生对大鼠脑缺血-再灌注性损伤起到保护作用,且该保护作用也可能与抑制神经细胞凋亡有关。

关 键 词:再灌注损伤  神经再生  细胞凋亡  大鼠  脑缺血-再灌注  络瘀通胶囊

Effect of Luoyutong capsule on focal cerebral ischemia-reperfusion injury in rats
Institution:WANG Li-ye, ZHAO Hai-ping, WANG Rong-liang, ZHANG Chen-cheng, LIU Xiao-meng, WANG Ning-qun, LUO Yu-min, CHEN Zhi-gang ( Department of Neurology (I), Dongfang Hospital, Beijing University of Chinese Medicine, Beijing 100078, China)
Abstract:Objectives To observe the effect of Luoyutong capsule on neurological function following focal cerebral ischemia-reperfusion in rats and to preliminarily study the protective mechanism of Luoyutong capsule for focal cerebral ischemia-reperfusion in rats. Methods A rat model of middle cerebral artery occlusion (MCAO)was induced by the modified Longa method. After 1. 5 h of ischemia,reperfusion started. Ten male SD rats were selected as sham operation group,and forty male SD rats were randomly divided into 4 groups:Model (MCAO),Luoyutong moderate-dose (LYTM),Luoyutong high-dose (LYTH),and citicoline sodium (CS)groups (n=10 in each group). At day 3 and 7 after modeling,the neurological function of the rats was evaluated by using 12 neurological score and forelimb placing test. Western blotting was used to detect the expression levels of brain derived neurotrophic factor (BDNF),basic fibroblast growth factor (b-FGF),and phosphor/protein kinase (p-AKT/AKT)on the ischemic side of the rats and in the ipsilateral brain tissue at day 3 after modeling,as well as the expression level of Caspase-12 at day 7 after modeling in the ipsilateral brain tissue,and a comparison was performed among the groups. Results (1 )Neurological score:At day 3 after modeling,there was no significant difference between the 12 neurological score and the forelimb placing test score (all P〉0. 05);At day 7 after modeling, there were obvious improvement in the LYTM,LYTH,and CS groups compared with model group (all P〈0.05). (2)The results of western blot showed that①compare with the sham operation group,the expression levels of BDNF and b-FGF were reduced obviously (all P〈0.05);compare with the MCAO group,the expression levels of the LYTM,LYTH and CS groups could be up-regulated,particularly in the LYTH group (P〈0. 01);② compare with the sham operation group,the expression level of p-AKT/AKT in MCAO group was decreased obviously (P〈0. 05);compare with the MCAO
Keywords:Reperfusion injury  Nerve regeneration  Apoptosis  Rats  Cerebral ischemia-reperfusion  Luoyutong capsule
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