KCNQ通道开放剂QO-83对6 Hz刺激癫痫模型小鼠的作用研究  

Effects of KCNQ Channel Opener QO-83 on 6 Hz Stimulated Epilepsy Model Mice

作　　者：王梓宇 王香雨 李金灿 吕添熠 李依萱 王梓橦 马梦妍 贾庆忠[1,2,3] WANG Ziyu;WANG Xiangyu;LI Jincan;LV Tianyi;LI Yixuan;WANG Zitong;MA Mengyan;JIA Qingzhong(School of Pharmacy,Hebei Medical University,Shijiazhuang 050017,China;Key Laboratory of Innovative Drug Research and Evaluation of Hebei Province,Shijiazhuang 050017,China;Key Laboratory of New Drug Pharmacology and Toxicology of Hebei Province,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学药学院,石家庄050017 [2]河北省新药非临床评价工程实验室,石家庄050017 [3]河北省药理毒理研究重点实验室,石家庄050017

出　　处：《中国药学杂志》2024年第22期2148-2158,共11页Chinese Pharmaceutical Journal

基　　金：河北省重点研发计划项目资助(21372601D);河北省自然科学基金项目资助(H2021206352);河北医科大学2023年大学生创新性实验计划项目资助(USIP2023200)。

摘　　要：目的通过建立并优化6 Hz点燃模型,探究KCNQ通道新型开放剂化合物QO-83抗癫痫活性,为QO-83在耐药性癫痫中的应用提供科学依据。方法将小鼠分为对照组、模型组、阳性药左乙拉西坦(100 mg·kg^(-1))组、对照药卡马西平(50 mg·kg^(-1))组和QO-83低(2 mg·kg^(-1))、中(4 mg·kg^(-1))、高(6 mg·kg^(-1))剂量组。建立6 Hz点燃癫痫模型小鼠,腹腔注射相应药物,30 min后再次施加6 Hz角膜刺激,进行癫痫行为学评价。随后进行一系列精神行为学实验:悬尾实验、T迷宫实验、新物体识别实验,分别观察小鼠的不动时间、选择正确率、认知指数,评价动物的抑郁倾向和学习记忆能力。对6 Hz点燃癫痫模型小鼠进行免疫荧光染色,观察c-Fos蛋白的表达变化,评价QO-83对特定脑区的保护作用;观察其靶点KCNQ2蛋白的表达变化,评价其对靶蛋白表达的影响。结果传统抗癫痫药卡马西平(50 mg·kg^(-1))对建立的模型小鼠无明显作用,阳性药物左乙拉西坦(100 mg·kg^(-1))有效,分别给予2、4、6 mg·kg^(-1)QO-83时,呈现剂量依赖性抗癫痫作用,可使点燃小鼠的癫痫大发作率及发作时间均显著下降。4 mg·kg^(-1)的QO-83可明显减少癫痫小鼠在悬尾实验中的不动时间,增加小鼠T迷宫正确率,增加小鼠新物体识别指数。QO-83可明显下调癫痫小鼠海马和梨状皮质区的c-Fos表达;上调晚期模型小鼠CA3、DG区周围的神经纤维和PIR区KCNQ2蛋白的表达。结论化合物QO-83具有良好的抗耐药性癫痫活性,其对癫痫动物的保护情况优于阳性药物左乙拉西坦,同时还具有改善癫痫动物精神行为的作用,表现出对耐药性癫痫的治疗潜力,其作用机制可能与上调靶点蛋白KCNQ2表达相关。OBJECTIVE To explore the antiepileptic activity of QO-83,a novel KCNQ channel opener,by establishing and optimizing a 6 Hz corneal kinked model,and to provide a scientific basis for the application of QO-83 in drug-resistant epilepsy.METHODS The mice were divided into control group,model group,positive drug levetiracetam(100 mg·kg^(-1))group,control drug carbamazepine(50 mg·kg^(-1))group,and QO-83 low(2 mg·kg^(-1)),medium(4 mg·kg^(-1)),and high(6 mg·kg^(-1))dose groups.The 6 Hz kindled mice were established,and drugs was injected intraperitoneally.After 30 min,6 Hz corneal stimulation was applied again to observe its protective effect on the animal.And set up a series of behavioral experiments:tail suspension test,T-maze test,and new object recognition test,observe the immobility time,selection accuracy,and recognition index of mice respectively,to evaluate its impact on animal depressive tendencies and memory abilities.Immunofluorescence staining was performed on 6 Hz model mice to observe the expression changes of c-Fos protein and evaluate the protective effect of QO-83 on specific brain regions.Observe the expression changes of its target KCNQ2 protein and evaluate its impact on target protein expression.RESULTS The established model mice were not sensitive to the action of carbamazepine(50 mg·kg^(-1)),but effective against levetiracetam(100 mg·kg^(-1)).When given 2,4 and 6 mg·kg^(-1) QO-83,dose-dependent anti-epileptic effects can be observed,which significantly reduced the incidence and duration of seizures in ignited mice.The 4 mg·kg^(-1) of QO-83 can significantly reduce the immobility time of epileptic mice in tail suspension experiments,increase the accuracy of mice in T-maze test,and increase the recognition index(RI)of mice in new object recognition test.QO-83 can significantly downregulate the expression of c-fos in the hippocampus and piriform cortex of epileptic mice,and upregulate the expression of KCNQ2 protein in the CA3,DG regions and their surrounding fibers and PIR regions of late-st

关 键 词：耐药性癫痫 6Hz点燃模型 钾通道开放剂 KCNQ2 

分 类 号：R966[医药卫生—药理学]