基于PI3K/Akt/Nrf2信号通路研究逍遥散拆方药队挥发油部位对脂多糖致抑郁样模型小鼠的作用及机制  

Effect and mechanism of volatile oil from disassembled prescription of Xiaoyao San on LPS-induced depressive-like model mice based on PI3K/Akt/Nrf2 signaling pathway

作　　者：谢志强 胡靖文 曾九僧 陈力[2,3] 谢红潇[1,2] 方洋 曾南 XIE Zhiqiang;HU Jingwen;ZENG Jiuseng;CHEN Li;XIE Hongxiao;FANG Yang;ZENG Nan(College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;State Key Laboratory of Southwestern Chinese Medicine Resources,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Department of Pharmacy,Clinical Medical College and the First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都中医药大学药学院,四川成都611137 [2]成都中医药大学西南特色中药资源国家重点实验室,四川成都611137 [3]成都医学院第一附属医院药剂科,四川成都610500

出　　处：《中草药》2024年第7期2283-2291,共9页Chinese Traditional and Herbal Drugs

基　　金：成都中医药大学西南特色中药资源国家重点实验室开放基金项目(2020XSGG002);成都中医药大学杏林学者项目(QJJJ2022012)。

摘　　要：目的采用脂多糖(lipopolysaccharide,LPS)诱导的小鼠抑郁样模型,探究逍遥散拆方药队(柴胡、当归、薄荷,CDB)挥发油部位的抗抑郁作用,及对磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶(protein kinase B,Akt)/核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)信号通路的影响。方法雄性C57BL/6J小鼠按体质量分层随机分为对照组、模型组、盐酸氟西汀(10 mg/kg)组和CDB挥发油高、低剂量(73.6、36.8 mg/kg)组。小鼠连续ig给药16 d,给药第13~15天,ip 1 mg/kg LPS(20 mL/kg)建立模型。通过糖水偏好测试、强迫游泳实验和悬尾实验观察动物抑郁样行为;ELISA法检测小鼠血清白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;化学法检测小鼠海马组织丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)水平和超氧化物歧化酶(superoxide dismutase,SOD)活性;尼氏染色法检测小鼠海马尼氏小体数量;免疫荧光法检测小鼠海马CA3区离子钙结合衔接分子-1(ionized calcium-binding adapter molecule-1,Iba-1)表达;Western blotting检测小鼠海马脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、p-PI3K/PI3K、p-Akt/Akt、Nrf2、血红素加氧酶-1(heme oxygenase-1,HO-1)、SOD1、p-p65/p65、IL-1β、Iba-1蛋白表达。结果CDB挥发油显著提高模型小鼠糖水偏好率(P<0.01),缩短强迫游泳和悬尾实验不动时间(P<0.05、0.01);显著降低海马MDA水平(P<0.05),提高GSH水平及SOD活性(P<0.05),降低血清IL-1β和TNF-α水平(P<0.01);显著增加小鼠海马CA3区尼氏小体平均荧光强度并抑制CA3区Iba-1表达(P<0.05、0.01);显著上调海马组织BDNF、p-PI3K/PI3K、p-Akt/Akt、Nrf2、HO-1和SOD1表达水平(P<0.05、0.01),抑制p-p65/p65、IL-1β、Iba-1蛋白表达(P<0.05、0.01)。结论CDB挥发油能激活PI3K/Akt/Nrf2信号通路,提高抗氧化水平并减轻神经炎症,从而发挥抗抑郁作用。Objective Using lipopolysaccharide(LPS)to induce a depression-like model in mice to investigate the antidepressant effects of volatile oil parts of disassembled prescription[Chaihu(Bupleuri Radix),Danggui(Angelicae Sinensis Radix),Bohe(Menthae Haplocalycis Herba),CDB]of Xiaoyao San(逍遥散)and its effects on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor erythroid 2-related factor 2(Nrf2)signaling pathway.Methods Male C57BL/6J mice were stratified by body weight and randomly divided into control group,model group,fluoxetine hydrochloride(10 mg/kg)group and CDB volatile oil highand low-dose(73.6,36.8 mg/kg)groups.Mice in each group were gavaged 16 consecutive days,and the model was established by intraperitoneal injection of 1 mg/kg LPS(20 mL/kg)on 13th to 15th day of administration.Depression-like behavior was observed by sucrose preference test,forced swimming test and tail suspension test.Levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in serum were measured by ELISA;Levels of malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD)activity in hippocampus of mice were measured by chemically;The number of mouse hippocampal Nissl bodies was detected by Nissen staining;Immunofluorescence method was used to detect the expression of ionized calcium-binding adapter molecule-1(Iba-1)in CA3 area of mouse hippocampus;Western blotting was used to detect brain-derived neurotrophic factor(BDNF),p-PI3K/PI3K,p-Akt/Akt,Nrf2,heme oxygenase-1(HO-1),SOD1,p-p65/p65,IL-1βand Iba-1 protein expressions in hippocampus.Results CDB volatile oil significantly increased the rate of sucrose preference in model mice(P<0.01),shortened the immobility time of forced swimming and tail suspension test(P<0.05,0.01),significantly decreased the hippocampal MDA level(P<0.05),increased GSH level and SOD activity(P<0.05),decreased the levels of IL-1βand TNF-αin serum(P<0.01),significantly increased the average fluorescence intensity of Nissl bodies in hippocampal CA3 region of mice and inh

关 键 词：逍遥散 挥发油 抑郁症 氧化应激 神经炎症 磷脂酰肌醇3-激酶/蛋白激酶/核因子E2相关因子2信号通路 藁本内酯 薄荷脑 香芹酮 丁烯基苯酞 

分 类 号：R285.5[医药卫生—中药学]