基于TLR4/NF-κB信号通路探讨益气通脉方对ApoE^(-/-)小鼠主动脉粥样硬化的作用机制  

Mechanism of Yiqi Tongmai Formula in Treatment of Atherosclerosis of Aorta in ApoE^(-/-)Mice Based on TLR4/NF-κB Signaling Pathway

作　　者：陈雨晗 任权娜 汪坤 杨艳华 马鸣 张雨 赵旭 CHEN Yu-han;REN Quan-na;WANG Kun;YANG Yan-hua;MA Ming;ZHANG Yu;ZHAO Xu(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou,450046;Department of Pharmacy,Henan Provincial Hospital of Traditional Chinese Medicine/The Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou,450002)

机构地区：[1]河南中医药大学药学院,郑州450000 [2]河南省中医院/河南中医药大学第二附属医院药学部,郑州450002

出　　处：《中国中西医结合杂志》2024年第7期835-842,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：河南省中医药科学研究专项(No.20-21ZYZD08,No.2023ZY1015,No.2023ZY2081);河南省中医药拔尖人才培养项目(No.豫卫中医函[2021]15号)。

摘　　要：目的 探讨益气通脉方调节Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路对载脂蛋白E^(-/-)(ApoE^(-/-))小鼠动脉粥样硬化(AS)炎症反应的作用机制。方法 将8只C57BL/6J小鼠作为空白组,40只同周龄雄性ApoE^(-/-)小鼠随机分为模型组、益气通脉方低、中、高剂量组[0.39、0.78、1.56g/(kg·d)]和阿托伐他汀组[2.6 mg/(kg·d)],每组8只。除空白组外采用高脂饲料诱导AS模型,各给药组分别给予相应药物灌胃,空白组与模型组给予等体积0.9%氯化钠溶液灌胃,连续12周。采用酶标仪检测小鼠血脂水平;ELISA检测小鼠血清中肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)水平;油红O染色观察小鼠主动脉斑块面积;HE、Masson染色观察小鼠主动脉窦内膜斑块病理变化;免疫组化(IHC)、免疫荧光(IF)法检测主动脉窦中TLR4、髓细胞分化初级反应蛋白88(MyD88)的表达;Western Blot检测小鼠主动脉中TLR4/NF-κB信号通路相关因子蛋白表达。结果 与空白组比较,模型组小鼠血清甘油三酯(TG)、总胆固醇(TC)及低密度脂蛋白胆固醇(LDL-C)水平升高(P<0.01),高密度脂蛋白胆固醇(HDL-C)水平下降(P<0.01);血清TNF-α、MCP-1含量升高(P<0.01);主动脉窦内膜斑块以及胶原面积增加(P<0.01),MyD88表达升高(P<0.01);主动脉TLR4、NF-κB、核因子κB抑制蛋白α(IκBα)、MyD88蛋白表达升高(P<0.01)。与模型组比较,益气通脉方中、高剂量组和阿托伐他汀组血清TC、TG、LDL-C水平下降(P<0.01),血清中TNF-α、MCP-1含量下降(P<0.01),主动脉窦斑块以及胶原面积均下降(P<0.01,P<0.05),主动脉窦MyD88水平表达下降(P<0.01),主动脉TLR4、NF-κB、MyD88、IκB蛋白表达下降(P<0.01,P<0.05)。结论 益气通脉方可改善血脂和炎症因子的水平,减少主动脉窦中斑块脂质堆积,其作用机制可能与下调TLR4/NF-κB信号通路有关。Objective To investigate the effect of Yiqi Tongmai Formula(YTF)on the inflammatory response of apolipoproteinE^(-/-)(ApoE^(-/-))mice atherosclerosis(AS)based on the regulation of Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signalling pathway.Methods Forty male ApoE^(-/-)mice were randomly divided into model group,the low,medium and high groups of YTF(0.39,0.78,1.56 g·kg^(-1)·d^(-1)),and the atorvastatin group(2.6 mg·kg^(-1)·d^(-1)),8 mice in each group,and 8 C57BL/6J mice of the same week of age as the blank control group.Except for the blank group,the AS model was induced by high fat diet.Each drug group was given the corresponding drug,and the blank group and model group were given the equal volume 0.9%sodium chloride solution for 12 consecutive weeks.The blood lipid level of mice was detected by enzyme marker assay;the expression of tumor necrosis factor-α(TNF-α)and monocyte chemotactic protein-1(MCP-1)in mouse serum were detected by ELISA;and the area of aortic plaque in mice was observed by oil red O staining;HE and Masson staining were used to observe the pathological changes of intima-media plaques in mouse aortic sinus;immunohistochemistry(IHC)and immunofluorescence(IF)methods were used to detect the expression of TLR4 and myeloid differentiation primary response protein88(MyD88)in aortic sinus;and Western Blot was used to detect the expression of TLR4/NF-κB signaling pathway-related factor protein in mouse aorta.Results Compared with the blank group,the serum levels of triglyceride(TG),total cholesterol(TC)and lowdensity lipoprotein cholesterol(LDL-C)significantly increased(P<0.01),high-density lipoprotein cholesterol(HDL-C)levels significantly decreased in the model group(P<0.01).Serum TNF-α,MCP-1 levels significantly increased(P<0.01).The intima-media plaque as well as collagen fibres area in aortic sinus increased(P<0.01),the protein expression of MyD88 increased(P<0.01).The protein expression of aortic arch TLR4,NF-κB,nuclear factorκB inhibitor alpha(IκBα),and MyD88 significantl

关 键 词：益气通脉方 动脉粥样硬化 炎症反应 TOLL样受体4 核转录因子-ΚB 中药复方 

分 类 号：R285.5[医药卫生—中药学]