基于AMPK/Nrf2信号通路探讨柴胡龙骨牡蛎加味方对癫痫大鼠能量代谢的干预机制  

Investigating themechanism of modified Chaihu Longgu Muli Decoction's intervention in energy metabolism in epileptic rats based on the AMPK/Nrf2signaling pathway

作　　者：韩慧[1] 张可颐 王颖[1] 张子环 李洪斌 HAN Hui;ZHANG Keyi;WANG Ying(Beichen Hospital of Traditional Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 300400,China;Graduate School,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津市北辰区中医医院/天津中医药大学附属北辰中医医院,天津300499 [2]天津中医药大学,天津301617

出　　处：《四川中医》2025年第4期143-148,共6页Journal of Sichuan of Traditional Chinese Medicine

基　　金：天津市卫健委天津市中医药管理局中医中西医结合科研课题(编号:2021124);天津市北辰区科技计划项目(编号:SHGY2023025)。

摘　　要：目的:探讨柴胡龙骨牡蛎加味方对癫痫大鼠能量代谢及腺苷酸活化蛋白激酶(AMPK)/核因子E2相关因子2(Nrf2)通路的影响。方法:采用戊四氮(PTZ)腹腔注射构建40只癫痫大鼠模型,随机分为空白组、模型组、中药组、西药组,每组各10只。均予治疗4周,模型组和空白组每日给予0.9%氯化钠灌胃,西药组给予丙戊酸钠缓释片灌胃,中药组给予汤剂柴胡龙骨牡蛎加味方灌胃。治疗28天后,尼氏染色观察各组大鼠海马组织神经元形态结构变化;Westernblot检测AMPK/Nrf2通路AMPKα、p-AMPKα及Nrf2蛋白含量变化与线粒体呼吸链复合体Ⅰ和复合体Ⅲ的蛋白表达变化。RT-PCR检测Nrf2mRNA的表达水平。结果:①各组大鼠海马组织神经元形态结构变化:与模型组相比,中药和西药组的海马组织损伤程度较轻,且中药组的神经元丢失和变性情况最不明显,尼氏小体数量与空白组最接近。②AMPK/Nrf2通路相关蛋白含量变化:模型组大鼠海马中AMPKα、p-AMPKα、ND1、UQCRC2以及Nrf2蛋白含量均显著降低。中药组和西药组AMPKα、p-AMPKα、ND1、UQCRC2以及Nrf2蛋白含量均提高(P<0.05),中药组在提升Nrf2蛋白含量方面效果更为显著。③RT-PCR检测:与空白组相比,模型组大鼠的海马中Nrf2 mRNA的表达水平显著降低。中药和西药均能有效提高Nrf2mRNA的表达水平(P<0.05)。结论:柴胡龙骨牡蛎加味方可能通过上调AMPKα1、Nrf2的表达量,激活AMPK/Nrf2通路,缓解氧化应激反应,调节线粒体能量代谢和修复癫痫大鼠海马神经元损伤。Objective:To investigate the effects of modified Chaihu Longgu Muli Decoction on energy metabolism and the AMP-activated protein kinase(AMPK)/nuclear factor erythroid 2-related factor 2(Nrf2)pathway in epileptic rats.Methods:A model of epilepsy was induced in 40 rats via intraperitoneal injection of pentylenetetrazol(PTZ).The rats were randomly divid-ed into four groups:blank group,model group,traditional Chinese medicine(TCM)group,and western medicine(WM)group,with 10 rats in each group.All groups were treated for four weeks.The model and blank groups received a daily intragastric admin-istration of 0.9%sodium chloride,the WM group received intragastric administration of sodium valproate sustained-release tab-lets,and the TCM group received intragastric administration of modified Chaihu Longgu Muli Decoction.After 28 days of treatment,Nissl staining was used to observe morphological changes in the hippocampal neurons of each group.Western blot analysis was used to detect changes in AMPKα,p-AMPKα,and Nrf2 protein levels in the AMPK/Nrf2 pathway,as well as changes in the protein expression of mitochondrial respiratory chain complexes I and III.RT-PCR was used to detect the expression levels of Nrf2 mRNA.Results:1.Hippocampal Neuronal Morphology:compared with the model group,the TCM and WM groups exhibited less hippocampal tissue damage,with the TCM group showing the least neuronal loss and degeneration,and the number of Nissl bodies being closest to the normal group.2.AMPK/Nrf2 Pathway Protein Levels:the model group showed significantly reduced levels of AMPKα,p-AMPKα,ND1,UQCRC2,and Nrf2 proteins in the hippocampus.Both the TCM and WM groups had in-creased levels of these proteins(P<0.05),with the TCM group showing a more significant effect in increasing Nrf2 protein lev-els.3.RT-PCR Detection:compared with the blank group,the model group had significantly lower expression levels of Nrf2 mR-NA in the hippocampus.Both the TCM and WM groups effectively increased the expression levels of Nrf2 mRNA(P<0.05).Conclus

关 键 词：柴胡龙骨牡蛎加味方 线粒体能量 癫痫 作用机制 

分 类 号：R285[医药卫生—中药学]