异鼠李素对ApoE^(-/-)小鼠动脉粥样硬化的保护作用及机制研究  

Isorhamnetin Ameliorates Pro-Atherosclerosis in Apolipoprotein E-Deficient Mice

作　　者：吴义林 江礼娟 罗义波 袁点 李为真[2] WU Yilin;JIANG Lijuan;LUO Yibo;YUAN Dian;LI Weizhen(Department of Cardiology,Jiading Branch of Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 201803,China;Department of Cardiology,Shanghai First People's Hospital,Shanghai 201803,China)

机构地区：[1]上海市第一人民医院嘉定医院/上海市嘉定区江桥医院心内科,上海201803 [2]上海市第一人民医院心内科,上海201803

出　　处：《转化医学杂志》2024年第4期632-637,共6页Translational Medicine Journal

基　　金：上海市嘉定区卫健委资助项目(2022-KY-03)。

摘　　要：目的探讨异鼠李素对ApoE^(-/-)小鼠动脉粥样硬化的保护作用及潜在分子机制。方法细胞实验中,巨噬细胞随机分为对照组、氧化低密度脂蛋白ox-LDL组及异鼠李素5、10、20μmol/L组,并予相应的干预措施。采用油红O染色法检测各组巨噬细胞内泡沫细胞占比,采用Western blot法检测巨噬细胞中Toll样受体4(TLR4)、CD36、磷酸化核因子κB抑制蛋白α/核因子κB抑制蛋白α(p-IκBα/IκBα)蛋白表达水平,流式细胞术检测各组巨噬细胞凋亡水平。动物实验中,选取6只6周龄雄性C57BL/6小鼠作为对照组,18只6周龄雄性ApoE-/-小鼠随机分为模型组及异鼠李素中、高剂量组。所有小鼠均采用高脂饲料喂养;异鼠李素中、高剂量组分别予以异鼠李素100、200 mg(/kg·d)灌胃给药。检测各组小鼠干预后主动脉组织相关细胞因子白细胞介素-1β(IL-1β)、IL-18及核转录因子κB p65(NF-κB p65)水平,氧化应激指标一氧化氮(NO)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平;采用实时荧光定量聚合酶链式反应(RT-qPCR)技术检测各组主动脉均浆组织IL-18、髓样分化因子88(MyD88)、基质金属蛋白酶13(MMP-13)、TLR4、NF-κB p65 mRNA表达水平;油红O染色观察各组小鼠主动脉粥样硬化情况。结果在细胞学实验中,异鼠李素5、10、20μmol/L组泡沫细胞占比均低于ox-LDL组(P<0.05);ox-LDL组CD36、TLR4、p-IκBa/IκBa蛋白表达水平及细胞凋亡水平均显著高于对照组(P<0.05);异鼠李素5、10、20μmol/L组CD36、TLR4、p-IκBa/IκBa蛋白表达水平及细胞凋亡水平均显著低于ox-LDL组(P<0.05)。在动物实验中,模型组小鼠IL-1β、IL-18、NF-κB p65、NO、MDA水平及TLR4、NF-κB p65、MyD88、IL-18 mRNA表达水平、油红O染色斑块面积均高于对照组(P<0.05),SOD浓度及MMP-13 mRNA表达水平均低于对照组(P<0.05);异鼠李素中、高剂量组小鼠IL-1β、IL-18、NF-κB p65、NO、MDA水平及TLR4、NF-κB p65、MyD88、IL-18 mRNA表达水Objective To explore the protective effect of isorhamnetin on atherosclerosis in ApoE^(-/-)mice and its potential molecular mechanisms.Method In the cell experiment,macrophages were randomly divided into control group,model group,and low,medium,and high concentration isorhamnetin intervention groups.Lipid content and foam cell proportion in macrophages were detected using oil red O staining.Western blot was used to measure the protein expression levels of TLR4,CD36,and p-IκBα/IκBαin macrophages.Annexin V/PI staining was used to assess apoptosis levels in macrophages of each group.In the animal experiment,6 C57BL/6 mice were selected as the control group,and 18 ApoE^(-/-)mice were randomly divided into model group and medium/high-dose isorhamnetin intervention groups.All mice were fed a highfat diet;the medium and high intervention groups were orally administered isorhamnetin at doses of 100 and 200 mg/kg/day,respectively.ELISA was used to measure levels of IL-1β,IL-18,NF-κB p65,and MCP-1 in aortic homogenates.Levels of NO,MDA,and SOD were determined using spectrophotometry.qRT-PCR was employed to assess TLR4 and NF-κB p65 mRNA expression levels in each group.Results In the cell experiment,the low,medium,and high-dose isorhamnetin intervention groups showed significantly lower lipid content,foam cell proportion,apoptosis,and protein expression levels of CD36,TLR4,and p-IκBα/IκBαcompared to the model group(P<0.05).In the animal model,after 8 weeks of intervention,the model group exhibited significantly higher levels of IL-1β,IL-18,NF-κB p65,MCP-1,oxidative stress markers(NO and MDA),and aortic plaque area,as well as increased TLR4 and NF-κB p65 mRNA expression compared to the control group(P<0.05).The medium and high-dose isorhamnetin groups showed significantly reduced levels of IL-1β,IL-18,NF-κB p65,MCP-1,oxidative stress markers,and plaque area,along with increased SOD levels compared to the model group(P<0.05).Conclusion Isorhamnetin can inhibit inflammation and the pro-atherosclerosis pathway i

关 键 词：异鼠李素 动脉粥样硬化 ApoE^(-/-)小鼠 氧化应激炎症 TOLL样受体4 NF-κB 

分 类 号：R543.5[医药卫生—心血管疾病]