莪术抗抑郁有效成分筛选及其调控Nrf2/GPX4/GSH通路的作用机制  

Screening of Antidepressant Active Components from Curcumae Rhizoma and Its Mechanism in Regulating Nrf2/GPX4/GSH Pathway

作　　者：宋永贵[1] 段德林 赖美茜子 刘亚丽[1,2] 艾志福 朱根华[1] 徐焕华[1] 郑琴 杨明[1,3] 苏丹[1] SONG Yonggui;DUAN Delin;LAI Meixizi;LIU Yali;AI Zhifu;ZHU Genhua;XU Huanhua;ZHENG Qin;YANG Ming;SU Dan(Key Laboratory of Depression Animal Model Based on Traditional Chinese Medicine(TCM)Syndrome of Jiangxi Province Administration of TCM,Key Research Office for Evaluation of TCM Efficacy(Prevention and Treatment of Mental Disorders and Brain Diseases)of Jiangxi Province Administration of TCM,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Key Laboratory of Pharmacodynamics and Safety Evaluation,Health Commission of Jiangxi Province,Nanchang Medical College,Nanchang 330006,China;Jiangxi Guxiang Jinyun Comprehensive Health Industry Co.Ltd.,Nanchang 330029,China)

机构地区：[1]江西中医药大学,江西省中医药管理局抑郁症中医证候动物模型重点研究室,江西省中医药管理局中药药效(防治精神障碍脑疾病)评价重点研究室,南昌330004 [2]南昌医学院,江西省卫生健康药效与安全性评价重点实验室,南昌330006 [3]江西古香今韵大健康产业有限公司,南昌330029

出　　处：《中国实验方剂学杂志》2025年第6期211-221,共11页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家中医药多学科交叉创新团队项目(ZYYCXTD-D-202207);江西省杰青需求牵引类项目(20224ACB218008);江西中医药大学科技创新团队项目(CXTD22008)。

摘　　要：目的:对莪术抗抑郁有效成分进行筛选、评价,并从抗氧化角度探索其调控核因子E2相关因子2(Nrf2)/谷胱甘肽(GSH)过氧化物酶4(GPX4)/GSH通路的作用机制。方法:采用1,1-二苯基-2-三硝基苯肼(DPPH)、2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除实验检测包括莪术醇、莪术酮、莪术二酮、莪术烯、莪术烯醇、莪术双环烯酮、去氢莪术二酮、异莪术烯醇、莪术呋喃二烯酮、莪术呋喃二烯、蓬莪术环氧酮在内的莪术11种特征成分的体外抗氧化活性。通过慢性不可预知应激(CUMS)建立果蝇抑郁模型,将W1118野生型黑腹雄性果蝇随机分为空白组、模型组、莪术醇组、莪术酮组、莪术二酮组、莪术烯组、莪术烯醇组、莪术双环烯酮组、去氢莪术二酮组、异莪术烯醇组、莪术呋喃二烯酮组、莪术呋喃二烯组、蓬莪术环氧酮组和氟西汀组(10μmol·L^(-1)),其中,莪术11种特征成分给药组剂量均为0.1 g·L^(-1),空白组和模型组给予等体积的溶媒。采用糖水偏好实验、攀爬实验、强迫游泳实验,评价果蝇抑郁行为学指标,采用液相色谱-质谱法评价果蝇大脑中5-羟色胺(5-HT)和多巴胺(DA)的水平,并采用熵权法综合评价确定莪术抗抑郁有效成分。此外,将7周龄C57BL/6J小鼠随机分为空白组,模型组,莪术烯低、高剂量组(0.5、1 mg·kg^(-1))和氟西汀组(10 mg·kg^(-1)),采用CUMS建立抑郁小鼠模型,结合行为学确认最佳活性成分莪术烯的抗抑郁效果。流式细胞术检测小鼠海马中活性氧(ROS)的含量;酶联免疫吸附测定法(ELISA)检测三磷酸腺苷(ATP)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、GSH含量的影响;透射电镜观察莪术烯对海马组织线粒体超微结构的影响;蛋白免疫印迹法(Western blot)测定莪术烯对Nrf2蛋白水平的影响,并采用Nrf2抑制剂ML385验证其抗抑郁作用与调控Nrf2的关系;实时荧光定量聚合酶链式反应Objective:To screen and evaluate the antidepressant compounds of Curcumae Rhizoma,and explore its mechanism of regulating the nuclear factor erythroid 2-related factor 2(Nrf2)/glutathione(GSH)peroxidase 4(GPX4)/GSH pathway from an antioxidant perspective.Methods:The antioxidant activities in vitro of 11 characteristic components from Curcumae Rhizoma,including curcumol,curgerenone,curdione,curzerene,curcumenol,curcumenone,dehydrocurdione,isocurcumenol,furanodienone,furanodiene and zederone,were detected using 1,1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid)diammonium salt(ABTS)radical scavenging assays.The depression in Drosophila melanogaster was induced by chronic unpredictable mild stress(CUMS),and W1118 wild-type male D.melanogaster were randomly divided into blank group,model group,curcumol group,curgerenone group,curdione group,curzerene group,curcumenol group,curcumenone group,dehydrocurdione group,isocurcumenol group,furanodienone group,furanodiene group,zederone group and fluoxetine group(10μmol·L^(-1)).The treatment groups received a dose of 0.1 g·L^(-1)of 11 characteristic components from Curcumae Rhizoma,while the blank and model groups were administered equivalent volumes of solvent.The sucrose preference test,climbing test and forced swimming test were used to evaluate the behavioral indicators of depression in D.melanogaster.Liquid chromatography-mass spectrometry(LC-MS)was used to detect the levels of 5-hydroxytryptamine(5-HT)and dopamine(DA)in the brain of D.melanogaster,and the entropy weight method was used to comprehensively evaluate neurobehavioral and neurotransmitter indicators,resulting in the identification of the antidepressant active components of Curcumae Rhizoma.In addition,a mouse depression model was established by CUMS,and C57BL/6J mice were randomly divided into blank group,model group,low and high dose groups of curzerene(0.5,1 mg·kg^(-1)),and fluoxetine group(10 mg·kg^(-1))to confirm the antidepressant effect of the optimal acti

关 键 词：莪术 抑郁症 药效评价 作用机制 氧化应激 核因子E2相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)/谷胱甘肽(GSH)通路 

分 类 号：R277[医药卫生—中医学] R749R971