失荅剌知丸及其拆方对局灶性脑缺血再灌注大鼠海马区ERK1/2通路的影响  

Effect of Shida Lazhi pills and its components on the ERK1/2 pathway in the hippocampus of rats with focal cerebral ischemia-reperfusion injury

作　　者：王娜琳 左艳丽 李占涛 王佩佩 贾戌生 冯佳婷 马玉玮 贾孟辉 WANG Nalin;ZUO Yanli;LI Zhantao;WANG Peipei;JIA Xusheng;FENG Jiating;MA Yuwei;JIA Menghui(School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia,China;The Second Affiliated Hospital of Ningxia Medical University,Yinchuan 750001,Ningxia,China;Key Laboratory of Hui Ethnic Medicine Modernization,Ministry of Education,Ningxia Medical University,Yinchuan 750004,Ningxia,China;Ningxia Chinese Medicine Research Center,Yinchuan 750021,Ningxia,China)

机构地区：[1]宁夏医科大学中医学院,宁夏银川750004 [2]宁夏医科大学第二附属医院,宁夏银川750001 [3]宁夏医科大学回医药现代化教育部重点实验室,宁夏银川750004 [4]宁夏回族自治区中医医院暨中医研究院,宁夏银川750021

出　　处：《湖南中医杂志》2020年第11期172-178,共7页Hunan Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81260568,81560816,81760906);宁夏回族自治区重点研发项目(2018BEG03078)。

摘　　要：目的:探讨失荅剌知丸及其拆方对脑缺血再灌注损伤模型大鼠的作用机制及其有效配伍成分。方法:将204只大鼠随机分为空白对照组(A组),假手术组(B组),模型组(C组),失荅剌知丸组(D组),巴豆组(E组),巴豆+三药组(F组),巴豆+余药组(G组),其中C、D、E、F、G组制备局灶性脑缺血再灌注模型,并按取材时间点不同分为1、3、7d组。各组大鼠取材前应用Garcia JH法进行神经功能评分;应用HE染色法观察大鼠各时间点脑组织病理学形态改变;免疫荧光法、Western Blot检测细胞外调节蛋白激酶1/2(ERK1/2)蛋白表达水平。结果:1)神经功能评分结果。与A、B组相比,其余各组评分均降低(P<0.01);与C组相比,D、E、F、G组评分均增加(P<0.01);各给药组中,D组评分高于E、F、G组,且E、F、G组中又以F组评分最高(P<0.05或P<0.01);C、D、E、F、G组评分均随给药时间延长而增加。2)大鼠脑组织海马区HE染色结果。C组大鼠海马CA1区神经细胞较A、B组排列松散、紊乱、模糊,胞核与胞质界限欠清晰;与C组相比,D、E、F、G组神经细胞形态改善明显,且以D组改善最为明显;E、F、G组中以F组改善明显。3)大鼠脑组织海马区免疫荧光结果。C组较A、B组大鼠脑组织海马区p-ERK1/2的阳性细胞计数增加(P<0.01);与C组相比,D、E、F、G组p-ERK1/2阳性细胞计数升高(P<0.01);各给药组中,D组计数最多,E、F、G组中以F组计数最多(P<0.05或P<0.01);C、D、E、F、G组p-ERK1/2的阳性细胞计数均随给药时间延长而升高。4)Western-bolt结果。与A、B组相比,其余各组大鼠脑组织海马区p-ERK1/2活化程度均升高(P<0.01);与C组相比,D、E、F、G组p-ERK1/2活化程度均升高(P<0.01);各给药组中C组p-ERK1/2活化程度均优于E、F、G组,E、F、G组中以F组p-ERK1/2的活化程度最显著(P<0.05或P<0.01);C、D、E、F组p-ERK1/2活化程度均随给药时间延长而升高。结论:D、E、F、G组均可改善大鼠脑缺血再灌�Objective:To investigate the mechanism of action of Shida Lazhi pills and its components on arat model of cerebral ischemia/reperfusion injury and its effective compatible components.Methods:A total of 204 rats were randomly into blank control group(group A),sham-operation group(group B),model group(group C),Shida Lazhi pills group(group D),croton group(group E),croton+three drugs group(group F),and croton+the other drugs group(group G).The rats in groups C,D,E,F,and G were used to establish a model of focal cerebral ischemia-reperfusion injury and were divided into 1-,3-,and 7-day groups according to the time point for sample collection.The Garcia JH method was used to determine neurological score before sample collection;HE staining was used to observe the pathomorphological changes of brain tissue at each time point;immunofluorescence assay and Western blot were used to measure the protein expression of extracellular signal-regulated kinase 1/2(ERK1/2).Results:Compared with groups A and B,all the other groups had a significant reduction in neurological score(P<0.01),and compared with group C,groups D,E,F,and G had a significant increase in neurological score(P<0.01);group D had a significantly higher neurological score than groups E,F,and G,among which group F had the highest neurological score(P<0.05 or P<0.01);in groups C,D,E,F,and G,the neurological score increased with the increase in the duration of drug administration.HE staining showed that compared with groups A and B,group C had loose,disordered,and unclear arrangement of hippocampal CA1 neurons,with unclear boundary between the nucleus and the cytoplasm;compared with group C,groups D,E,F,and G had significant improvement in the morphology of neural cells,with the most significant improvement in group D;among groups E,F,and G,group F had significantly better improvement.Immunofluorescence assay of the hippocampus showed that compared with groups A and B,group C had a significant increase in the number of p-ERK1/2-positive cells in the hippocampus(P<0.

关 键 词：脑缺血再灌注 失荅剌知丸 拆方 ERK1/2 实验研究 

分 类 号：R285.5[医药卫生—中药学]