大补肝汤通过调控ROS/Nrf2/HO-1通路对广泛性焦虑症大鼠的改善作用  

Effects of Dabugan Decoction on the improvement of generalized anxiety disorder in rats via ROS/Nrf2/HO⁃1 pathway

作　　者：黄仕琦 谭国伟 李艳明 安耀荣[1,2,3] HUANG Shi-qi;TAN Guo-wei;LI Yan-ming;AN Yao-rong(Gansu University of Traditional Chinese Medicine,Lanzhou 730000,China;Gansu Key Laboratory for Traditional Chinese Medicine Excavation and Innovative Transformation,Lanzhou 730000,China;Gansu Engineering Laboratory for Creation and Manufacturing of New Traditional Chinese Medicine Products,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学,甘肃兰州730000 [2]甘肃省中医方药挖掘与创新转化重点实验室,甘肃兰州730000 [3]甘肃省中药新产品创制工程实验室,甘肃兰州730000

出　　处：《中成药》2024年第7期2202-2210,共9页Chinese Traditional Patent Medicine

基　　金：甘肃省自然科学基金项目(22JR1RA21);2022年度兰州市科技发展指导性计划项目(2022-5-164)。

摘　　要：目的探讨大补肝汤对广泛性焦虑症(GAD)模型大鼠的作用机制。方法大鼠采用慢性不可预知温和应激法(CUMS)造模21d,随机分为模型组、帕罗西汀组(1.8mg/kg)和大补肝汤高、中、低剂量组(13.2、6.6、3.3g/kg),另设空白组,各组给予相应药物灌胃,持续28d。干预结束后,检测行为学以分析大鼠焦虑状态,HE染色和尼氏染色观察海马组织病理变化及尼氏体变化,免疫荧光法检测脑组织ROS活性,试剂盒检测海马组织SOD、CAT活性和MDA、GSH水平,RT-qPCR法和Westernblot法检测海马组织Nrf2、Keap1、HO-1、NQO1、BDNF、5-HT1A、caspase-3、p62mRNA和蛋白表达。结果与模型组比较,大补肝汤各剂量组大鼠焦虑行为得到改善,大鼠在高架十字迷宫中开臂端运动距离及总距离、进入明箱次数及停留时间、进入旷场中央时间及次数增加(P<0.05,P<0.01),海马神经细胞数量增加,排列紧密,细胞体损坏改善,细胞中尼氏体增加,海马组织中ROS活性和MDA水平降低(P<0.05,P<0.01),SOD、CAT活性和GSH水平均升高(P<0.05,P<0.01),海马组织Nrf2、HO-1、NQO1、BDNF、5-HT1AmRNA及蛋白表达均升高(P<0.05,P<0.01),p62、Keap1、caspase-3mRNA及蛋白表达均降低(P<0.05,P<0.01)。结论大补肝汤可有效改善GAD大鼠焦虑状态,其机制可能与调控ROS/Nrf2/HO-1信号通路、改善海马神经元氧化应激损伤有关。AIM To explore the mechanism of Dabugan Decoction’s action on a rat model of generalized anxiety disorder(GAD).METHODS The rat models established by 21 days exposure to chronic unpredictable mild stress(CUMS)were then randomly divided into the model group,the paroxetine group(1.8 mg/kg)and the high-dose,medium-dose and low-dose Dabugan Decoction groups(13.2,6.6 and 3.3 g/kg),in contrast to the other normal rats assigned into the blank group,for the following 28 days administration of the corresponding drugs by gavage.After the intervention,the rats had their anxiety intensity analyzed by behavioral tests;their hippocampal pathological changes and Nissl’s body differences observed by HE staining and Nissl staining;theircerebral ROS activity detected by immunofluorescence;their hippocampal activities of SOD and CAT,and levels of MDA and GSH detected by kit;and their hippocampal mRNA and protein expressions of Nrf2,Keap1,HO-1,NQO1,BDNF,5-HT1A,caspase-3 and GSH detected by RT-qPCR and Western blot.RESULTS Compared with the model group,the Dabugan Decoction groups displayed improved anxiety responses;increased movement distance at the open arm end and total distance as well,more entries and duration at the open box,more time and entries at the open field center in elevated plus maze(P<0.05,P<0.01);increased number of hippocampal neurons,ameliorated cell body damage,increased Nissl bodies in cells,decreased hippocampal ROS activity and MDA level(P<0.05,P<0.01);increased activities of SOD and CAT,and GSH level(P<0.05,P<0.01);increased hippocampal mRNA and protein expressions of Nrf2,HO-1,NQO1,BDNF and 5-HT1A(P<0.05,P<0.01),and decreased mRNA and protein expressions of p62,Keap1 and caspase-3(P<0.05,P<0.01).CONCLUSION Dabugan Decoction can effectively improve the anxiety of GAD rats,and its mechanism may be related to the recovery of oxidative stress-induced injury in hippocampal neurons via ROS/Nrf2/HO-1 signaling pathway.

关 键 词：大补肝汤 广泛性焦虑症 氧化应激 ROS/Nrf2/HO-1信号通路 海马 

分 类 号：R285.5[医药卫生—中药学]