基于Nrf2/Keap1通路探讨柴贝止痫汤对海人酸致痫大鼠的神经保护作用机制  

Neuroprotective mechanism of Chaibei Zhixian decoction for rats with kainicacid-induced epilepsy based on Nrf2/Keap1 signal pathway

作　　者：徐薇薇 赵颖[2] 王苏妹[2] 王越[3] 王莹[1] 张宓 杨松仪 刘金民[2] XU Weiwei;ZHAO Ying;WANG Sumei;WANG Yue;WANG Ying;ZHANG Mi;YANG Songyi;LIU Jinmin(Beijing University of Chinese Medicine,Beijing 100029,China;Dongfang Hospital Affiliated to Beijing University of Chinese Medicine,Beijing 100078,China;Tianjin University of Chinese Medicine,Tianjin 301617,China)

机构地区：[1]北京中医药大学,北京100029 [2]北京中医药大学东方医院,北京100078 [3]天津中医药大学,天津301617

出　　处：《现代中西医结合杂志》2023年第9期1169-1175,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：中央高校基本科研业务费专项资金资助项目(2020-JYB-ZDGG-133);国家自然科学基金面上项目(81774277);天津市自然科学基金项目(2019KJ049)。

摘　　要：目的探讨柴贝止痫汤对海人酸致痫大鼠氧化应激反应及核因子E2相关因子2(Nrf2)/Kelch样ECH相关蛋白1(Keap1)信号通路的影响。方法雄性Wistar大鼠30只,随机取10只作为假手术组,余20只采用海人酸侧脑室注射方法建立慢性复发性癫痫模型。将造模成功的大鼠随机分为癫痫组及柴贝止痫汤组各8只,柴贝止痫汤组大鼠给予柴贝止痫汤8.48 g/kg灌胃,假手术组和癫痫组大鼠给予生理盐水灌胃,均1次/d,连续8周。采用Morris水迷宫行为学实验评估大鼠空间学习及记忆能力,试剂盒检测血清丙二醛(MDA)含量和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,HE染色观察海马组织病理形态,Western blot法检测海马组织中Nrf2及Keap1蛋白表达情况。结果与假手术组比较,癫痫组大鼠定位航行实验逃避潜伏期明显延长(P<0.05);空间探索实验中总路程明显延长(P<0.05),平台区域路程及平台象限停留时间均明显缩短(P均<0.05),穿越平台次数明显减少(P<0.05);血清MDA含量明显升高(P<0.05),血清SOD及CAT活性均明显降低(P均<0.05);HE染色显示大鼠脑组织结构重度异常,海马CA1、CA3及DG区可见大量神经元变性,固缩深染,神经元纤维缠结;海马组织中Keap1蛋白表达量明显增加(P<0.05),Nrf2蛋白表达量明显降低(P<0.05)。与癫痫组比较,柴贝止痫汤组大鼠逃避潜伏期和总路程均明显缩短(P均<0.05),平台区域路程及平台象限停留时间均明显延长(P均<0.05),穿越平台次数明显增加(P<0.05);血清MDA含量明显降低(P<0.05),SOD及CAT活性均明显增高(P均<0.05);HE染色显示大鼠脑组织结构轻度异常,可见少量神经元变性,固缩深染,个别神经元纤维缠结现象;海马组织中Keap1蛋白表达量明显降低(P<0.05),Nrf2蛋白表达量明显增加(P<0.05)。结论柴贝止痫汤可改善海人酸致痫大鼠空间学习及记忆能力,保护神经元细胞,作用机制可能与激活Nrf2/Keap1信号通路,抑制氧化应�Objective It is to explore the effect of Chaibei Zhixian decoction on the oxidative stress response and nuclear factor E2 related factor 2(Nrf2)/Kelch-like epichlorohydrin-associated protein1(Keap1)signal pathway in rats with kainic acid-induced epilepsy.Methods Thirty male Wistar rats were selected,in which 10 ones were randomly selected as the sham operation group,and the remaining 20 rats were used to establish chronic recurrent epilepsy models by lateral ventricular injection of kainic acid.The successfully modeled rats were randomly divided into epilepsy group and Chaibei Zhixian decoction group,with 8 rats in each group.The rats in the Chaibei Zhixian decoction group were given Chaibei Zhixian decoction 8.48 g/kg by gavage,and the rats in the sham operation group and epilepsy group were given normal saline by gavage,all once daily,continuously treated for 8 weeks.The spatial learning and memory ability of the rats in each group were assessed by Morris water maze behavioral experiments,the content of serum malonaldehyde(MDA)and the activities of superoxide dismutase(SOD)and catalase(CAT)were measured by ELISA and WST-8 kit,the pathological morphology of hippocampal tissues was observed by HE staining,and the expressions of Nrf2 and Keap1 protein in hippocampal tissues were detected by Western blot.Results Compared with the sham operation group,the escape latency of the rats in the epileptic group was significantly prolonged in the orientation test(P<0.05);the total distance was significantly increased(P<0.05),the traveled distance and the staying time in the platform quadrant were significantly decreased(both P<0.05),and the times of crossing the platform was significantly reduced in the spatial exploration test(P<0.05);the level of serum MDA was significantly increased(P<0.05),and the activities of serum SOD and CAT were significantly reduced(both P<0.05);HE staining showed severe structural abnormalities in the brain tissue of the rats,with a large number of neuronal degeneration,deep staining of solidific

关 键 词：癫痫 氧化应激 柴贝止痫汤 核因子E2相关因子2 Kelch样ECH相关蛋白1 

分 类 号：R-332[医药卫生]