基于定痫汤干预后癫痫小鼠海马区miRNA-204表达水平的变化及脑电图变化的研究  

Effects of Dingxian Decoction on Expression of miRNA-204 in Hippocampus and EEG Changes in Epilepsy Mice

作　　者：卢玲 刁丽梅[2] 李欢 廖现秋 LU Ling;DIAO Limei;LI Huan;LIAO Xianqiu(Guangxi University of Chinese Medicine,Nanning 530001,China;The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《中医药学报》2022年第2期33-38,共6页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(81760809,81960858);广西中医药大学研究生教育创新计划项目(YCSY20190062);广西高等学校高水平创新团队及卓越学者计划(桂教人才[2020]6号);广西自然科学基金项目(2017GXNSFAA198294);广西卫生适宜技术研究与开发项(s2017049)。

摘　　要：目的:研究定痫汤干预后癫痫小鼠脑电图的变化、小鼠海马区miRNA-204表达水平的变化以及自噬相关蛋白ATG7、LC3II的表达变化,探讨其可能的作用机制。方法:取50只雄性昆明小鼠,其中40只随机分成模型组、生理盐水组、卡马西平组和定痫汤组,每组10只,另外10只为备用小鼠。首先对各组实验动物进行灌胃给药两周,模型组和生理盐水组均予20 mL·kg^(-1)·d^(-1)生理盐水进行灌胃;定痫汤组予7 g·kg^(-1)·d^(-1)定痫汤药液进行灌胃;卡马西平组予30 mg·kg^(-1)·d^(-1)卡马西平混悬液灌胃。然后,通过盐酸匹罗卡品构建小鼠急性癫痫模型。其次,对各组小鼠进行行为学和脑电图观察。最后,将小鼠断头取脑,采用实时荧光定量PCR技术检测小鼠海马区miRNA-204表达水平的变化;并且通过免疫组化法检测自噬相关蛋白ATG7、LC3II的表达变化。结果:与生理盐水组相比,模型组小鼠癫痫发作的频率多、持续时间长,小鼠脑电图尖波、棘波、棘慢波明显增多,海马区miRNA-204的表达明显下调(P<0.01),自噬相关蛋白ATG7、LC3II的表达明显增多(P<0.01)。与模型组相比,使用定痫汤干预后小鼠癫痫发作频率减少,小鼠脑电图痫性波形减少,小鼠海马区miRNA-204的表达升高(P<0.01),自噬相关蛋白ATG7、LC3II的表达下调(P<0.01)。结论:定痫汤可以减少小鼠癫痫发作的频率,减少小鼠脑电图的痫性波形。其可能是通过上调miRNA-204的表达,抑制自噬相关蛋白ATG7、LC3II的表达,拮抗小鼠海马神经元过度自噬,从而发挥抗癫痫的作用。Objective:To study the changes of EEG,the expression of miRNA-204 and the expression of autophagy related proteins of ATG7 and LC3 II in epilepsy mice after Dingxian Decoction intervention,thus to explore its possible action mechanism.Methods:40 male Kunming mice were randomly divided into the model group(group A),the normal saline group(group B),the Carbamazepine group(group C)and the Dingxian Decoction group(group D),with 10 mice in each group.Another 10 mice were taken as the spared.The experimental mice in each group were given intragastric administration for two weeks.The model group and the normal saline group were given normal saline at the concentration of 20 mL·kg^(-1)·d^(-1),the Dingxian Decoction group was treated with Dingxian Decoction at the concentration of 7 g·kg^(-1)·d^(-1),and the Carbamazepine group was treated with Carbamazepine Suspension at the concentration of 30 mg·kg^(-1)·d^(-1).Pilocarpine Hydrochloride was used to establish the epileptic model in mice.Mice behavior and EEG changes were observed in each group.The expression of miRNA-204 in hippocampus was detected by real-time quantitative PCR,and the expressions of autophagy related proteins of TG7 and LC3 II were detected by immunohistochemistry.Results:Compared to those in the normal saline group,the frequency and duration of epileptic seizures were the most and the longest,and the spike wave,spinous wave and slow wave of EEG were significantly increased,and the expression of miRNA-204 in hippocampus was significantly down-regulated in the model group(P<0.01);the expressions of autophagy related proteins of ATG7 and LC3 II increased significantly in the model group(P<0.01).Compared to those in the model group,the frequency of epileptic seizure was decreased,the epileptic waveform of EEG was reduced,the expression of miRNA-204 in hippocampus was increased(P<0.01),and the expressions of autophagy related proteins of ATG7 and LC3 II were down-regulated in the Dingxian Decoction group(P<0.01).Conclusion:Dingxian Decoction can reduce

关 键 词：定痫汤 癫痫 miRNA-204 脑电图 ATG7 LC3II 

分 类 号：R285.5[医药卫生—中药学]