摘要
目的研究对药“远志-天麻”对癫痫小鼠癫痫发作潜伏时间、空间记忆和情绪反应、ABAT/GAT1/GAD 65/GABRA1蛋白表达水平的影响,神经元的保护情况,探究该对药治疗癫痫的可能机制。方法将造模成功的小鼠随机分为模型组(n=6)、“远志-天麻”高剂量(YTH,300 mg·kg-1,n=6)、中剂量(YTM,100 mg·kg-1,n=6)、低剂量(YTL,30 mg·kg-1,n=6)组及对照组。采用旷场和高架十字检测对药“远志-天麻”对PTZ诱导的慢性癫痫小鼠模型空间记忆障碍和情绪反应的影响,HE染色观察小鼠脑组织海马区病变,以蛋白免疫印迹法检测小鼠脑海马区ABAT/GAT1/GAD 65/GABRA1蛋白表达水平并用免疫荧光检测小鼠脑组织中相关细胞免疫阳性数,分子模拟对接分析“远志-天麻”内标性成分与潜在靶标的作用情况。结果与模型组相比,对药“远志-天麻”能够显著延长小鼠癫痫发作的潜伏时间,改善戊四唑(PTZ)诱导的慢性癫痫小鼠的空间记忆障碍和情绪反应,调节GABA-AT/GAT-1分子信号通路的水平,下调脑海马内因癫痫发作所致的GAD 65、GABRA1、GABA-AT、GAT-1表达上升(P<0.05),减少海马神经元脱失,其内标性成分能够与GABA-AT及GABAA受体(GABA AR)较好结合。结论对药“远志-天麻”可以通过GABA-AT/GAT-1通路治疗癫痫,改善癫痫模型小鼠的空间记忆和情绪反应损伤,对神经元起到保护作用。
Objective To investigate the effects of herb pairs Polygala tenuifolia and Gastrodia elata on the latency,spatial memory,emotional response,ABAT/GAT1/GAD 65/GABRA1 protein expression levels,neuronal protection,and explore the possible mechanism of this drug in treating epilepsy in epileptic mice.Methods The successfully modeled mice were randomly divided into a model group(n=6),a high-dose(YTH,300 mg·kg-1,n=6),a medium dose(YTM,100 mg·kg-1,n=6),a low-dose(YTL,30 mg·kg-1,n=6)group of herb pairs Polygala tenuifolia and Gastrodia elata,and a control group.The effects of herb pairs Polygala tenuifolia and Gastrodia elata on spatial memory impairment and emotional response in PTZ induced chronic epilepsy mouse models were investigated using open field and elevated cross detection.HE staining was used to observe hippocampal lesions in mouse brain tissue,and protein immunoblotting was used to detect ABAT/GAT1/GAD 65/GABRA1 protein expression levels in mouse hippocampus.Immunofluorescence was used to detect the number of immune positive cells in mouse brain tissue.Molecular simulation docking analysis was performed to investigate the interaction between internal standard components of herb pairs Polygala tenuifolia and Gastrodia elata and potential targets.Results Compared with the model group,herb pairs Polygala tenuifolia and Gastrodia elata significantly prolonged the latency of epileptic seizures in mice,improved spatial memory impairment and emotional response induced by pentylenetetrazole(PTZ)in chronic epileptic mice,regulated the level of GABA-AT/GAT-1 molecular signaling pathway,downregulated the increased expression of GAD 65,GABRA1,GABA-AT,and GAT-1 in the hippocampus caused by epileptic seizures(P<0.05),and reduced hippocampal neuronal loss.Its internal standard components can bind well to GABA-AT and GABAA receptors(GABAAR).Conclusion Herb pairs Polygala tenuifolia and Gastrodia elata can treat epilepsy through the GABA-AT/GAT-1 pathway,improve spatial memory and emotional response damage in epilepsy model mice,and protect neurons.
作者
赵泽丰
年梦
王咏琪
吕红
姬晓彤
杨晓航
乔海法
ZHAO Zefeng;NIAN Meng;WANG Yongqi;LV Hong;JI Xiaotong;YANG Xiaohang;QIAO Haifa(Shaanxi Provincial Key Laboratory of Combination of Acupuncture and Medicine,Shaanxi Xianyang 712046,China;College of Acupuncture and Moxibustion,Shaanxi University of Chinese Medicine,Shaanxi Xianyang 712046,China)
出处
《陕西中医药大学学报》
2025年第1期22-32,共11页
Journal of Shaanxi University of Chinese Medicine
基金
国家自然科学基金项目(8210142829)
陕西省科技厅中医诊疗装备研发相关研究创新团队项目(303-202210705)
陕西省教育厅科研计划项目(21JS020)
陕西中医药大学经脉-脏腑相关研究创新团队项目(YL-09)。
关键词
远志
天麻
对药
癫痫
GABA-AT
GAT-1
Polygala tenuifolia
Gastrodia elata Blume
Drug pair
Epilepsy
GABA-AT
GAT-1
