基于Nrf2/GPX4通路调控铁死亡探讨黄连解毒汤对动脉粥样硬化小鼠的影响  

Effect of Huanglian Jiedutang in Regulating Ferroptosis in Mice with Atherosclerosis Based on Nrf2/GPX4 Signaling Pathway

作　　者：龚兆会[1,2] 高黎 翟惠奇 余锦紫 褚庆民[1,2] 罗川晋[1,2] 卿立金[1,2] 吴伟[1,2] 李荣[1,2] GONG Zhaohui;GAO Li;ZHAI Huiqi;YU Jinzi;CHU Qingmin;LUO Chuanjin;QING Lijin;WU Wei;LI Rong(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;The First Clinical School of Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;School of Traditional Chinese Medicine,Jinan University,Guangzhou 510632,China)

机构地区：[1]广州中医药大学第一附属医院,广州510405 [2]广州中医药大学第一临床医学院,广州510405 [3]暨南大学中医学院,广州510632

出　　处：《中国实验方剂学杂志》2025年第3期22-28,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省基础与应用基础研究基金项目(2022A1515110789,2022A1515220138,2023A1515220158,2022A1515220067);国家自然科学基金项目(82230126,82105047);广州中医药大学第一附属医院中青年骨干人才培育项目;广州中医药大学青年拔尖人才(团队)培育“揭榜挂帅”项目(A1-2601-24-414-110Z77);广州市科技计划项目(2023A04J1928)。

摘　　要：目的:研究黄连解毒汤通过改善铁死亡治疗动脉粥样硬化(AS)小鼠的作用机制。方法:取SPF级C57BL/6J小鼠10只为正常组,另取载脂蛋白E敲除(ApoE^(-/-))小鼠50只随机分为5组,分别为模型组、黄连解毒汤低、中、高剂量组和阿托伐他汀组(ATV组)。ApoE^(-/-)小鼠采用高脂饲料喂食8周构建AS模型,并在第9周开始分别予生理盐水,黄连解毒汤低、中、高剂量(3.9、7.8、15.6 g·kg^(-1)·d^(-1))和阿托伐他汀钙片(0.01 g·kg^(-1)·d^(-1))灌胃,共给药8周。采用大体油红O染色和马松(Masson)染色观察小鼠主动脉斑块的形成情况,自动生化分析仪测定血脂四项总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)水平,透射电镜观察主动脉线粒体结构,酶联免疫吸附测定法(ELISA)检测血清中超氧化物歧化酶(SOD)水平,微板法检测血清中还原型谷胱甘肽(GSH)含量,TBA法检测血清中丙二醛(MDA)含量,蛋白免疫印迹法检测小鼠主动脉核因子E_(2)相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)信号通路蛋白表达。结果:与正常组比较,模型组主动脉管腔斑块沉积,血清TC、LDL-C、TG、HDL-C、MDA含量显著升高(P<0.01),血清SOD、GSH和主动脉Nrf2、溶质载体家族7成员11(SLC7A11)、GPX4的表达水平均显著降低(P<0.01),主动脉线粒体碎裂、空泡化、体积萎缩,线粒体内嵴减少或者呈现松散、紊乱的形态。与模型组比较,黄连解毒汤低、中、高剂量组和ATV组主动脉管腔斑块沉积明显减少,小鼠血清TC、LDL-C、TG和MDA含量明显降低(P<0.05,P<0.01),血清SOD、GSH水平和主动脉Nrf2、SLC7A11、GPX4的表达水平升高(P<0.05,P<0.01),主动脉线粒体空泡化症状减轻,嵴数量增多且排序整齐。结论:黄连解毒汤能减轻AS小鼠主动脉管腔斑块沉积,降低血脂和MDA表达,升高SOD和GSH表达,改善铁死亡病理改变,其作用机制与Nrf2/GPX4信号通路有关。Objective:To study the mechanism of Huanglian Jiedutang(HLJDT)in treating mice with atherosclerosis(AS)by improving ferroptosis.Methods:A total of 10 SPF C57BL/6J mice were selected as a normal group,and 50 ApoE^(-/-)mice were randomly divided into five groups:model group,low-dose group of HLJDT,medium-dose group of HLJDT,high-dose group of HLJDT,and atorvastatin(ATV)group.ApoE^(-/-)mice were fed a high-fat diet for eight weeks to establish the AS model,and at the 9th week,they were given normal saline,low,medium,and high doses of HLJDT(3.9,7.8,15.6 g·kg^(-1)·d^(-1)),and atorvastatin calcium tablets(0.01 g·kg^(-1)·d^(-1)),respectively,for a total of eight weeks.The formation of aortic plaque in mice was observed by gross oil red O staining and Masson staining.The levels of total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),triglyceride(TG),and high-density lipoprotein cholesterol(HDL-C)in blood fat were measured by the automatic biochemical analyzer,and the mitochondrial structure of the aorta was observed by transmission electron microscopy.The content of serum superoxide dismutase(SOD)in serum was detected by enzyme-linked immunosorbent assay(ELISA).The content of reduced glutathione(GSH)in serum was detected by the microplate method,and that of malondialdehyde(MDA)in serum was detected by the TBA method.The protein expression of nuclear factor E_(2)-associated factor 2(Nrf2)/glutathione peroxidase 4(GPX4)signaling pathway was detected by Western blot.Results:Compared with those of the normal group,the contents of TC,LDL-C,TG,HDL-C,and MDA in the serum and the aortic vascular plaque deposition of the model group were significantly increased(P<0.01),while the expression levels of SOD and GSH in serum,as well as Nrf2,solute carrier family 7 member 11(SLC7A11),and GPX4 in aorta were significantly decreased(P<0.01).Mice in the model group appeared mitochondrial fragmentation and vacuolation in the aorta,volume atrophy,mitochondrial crista reduction,or a loose and disorganized form.Compared with th

关 键 词：黄连解毒汤 动脉粥样硬化 铁死亡 核因子E_(2)相关因子2(Nrf2)/谷胱甘肽过氧化物酶4(GPX4)信号通路 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R259