摘要
目的:体外模拟高脂血症小鼠心肌梗死(myocardial infarction,MI)和肾素-血管紧张素系统(RAS)被激活的内环境,研究氧化低密度脂蛋白(ox-LDL)能否在血管紧张素Ⅱ(AngⅡ)的基础上进一步诱导树突状细胞(dendritic cells,DCs)的成熟、迁移和炎症反应,并探讨可能参与其中的信号通路。方法:诱导C57BL/6J小鼠骨髓细胞分化成DCs,然后加入坏死心肌细胞上清,分别给予AngⅡ和ox-LDL+AngⅡ干预48h。流式细胞仪检测DCs的成熟标志物(CD83)和协同共刺激分子(CD86)的表达,用qPCR、ELISA方法测定炎性因子的表达。Western印迹法检测核因子κB(NF-κB)和IκB磷酸化程度,以及血凝素样氧化型低密度脂蛋白受体-1(LOX-1)和Toll样受体(TLR)4信号通路的表达。结果:ox-LDL能够在AngⅡ基础上进一步诱导CD83和CD86的表达,增强炎性因子和趋化因子的表达,增强NF-κB和IκB磷酸化程度(P<0.05)。ox-LDL能够在AngⅡ基础上进一步上调LOX-1的表达,同时激活了TLR4-MyD88-NF-κB通路(P<0.05)。结论:ox-LDL可能在AngⅡ的基础上进一步诱导DCs免疫成熟、迁移和炎症反应;LOX-1-TLR4-MyD88-NF-κB信号通路可能参与DCs成熟,以及DCs在高脂血症合并心肌梗死发生和发展中引起炎症反应的过程。
Objective：To explore the induction of maturation,migration,and inflammation of dendritic cells（DCs）after treatment with angiotensin（Ang）Ⅱ via oxidized low-density lipoprotein（ox-LDL）,and its corresponding signaling pathway.Methods：Bone marrow cells from C57 BL/6 J WT mice were differentiated into DCs.After adding supernatant of necrotic cardiomyocyte cells,DCs were treated with either AngⅡalone or in combination with ox-LDL for 48 h.CD83 and CD86 were measured via flow cytometry,while cytokines and chemokine levels were measured by the qPCR or ELISA.Nuclear factorκB（NF-κB）and IκB phosphorylation were detected by Western blotting.The expressions of lectin-like oxidized low-density lipoprotein receptor-1（LOX-1）,myeloid differentiation factor 88（MyD88）,and Toll-like receptor（TLR）4 were detected by Western blotting.Results：Ox-LDL could further up-regulate the maturity marker CD83 and CD86 in DCs on the basis of AngⅡtreatment.The expressions of inflammatory factors and chemokines,as well as phosphorylation of NF-κB and IκB were upregulated（P〈0.05）.The expression of LOX-1,phosphorylation of interleukin-1 receptor associated kinase（IRAK）-4 and the activation of TLR4/MyD88 signaling pathway were further enhanced by ox-LDL（P〈0.05）.Conclusions：The immune maturation,migration,and inflammation of DCs treated with AngⅡcould be further induced by ox-LDL,possibly viathe LOX-1-TLR4-MyD88 signaling pathway.
作者
马元吉
邹云增
葛均波
MA Yuan-ji;ZOU Yun-zeng;GE Jun-bo(Department of Cardiology,Zhongshan Hospital,Fudan University,Shanghai 200032,China)
出处
《中国临床医学》
2018年第3期383-386,共4页
Chinese Journal of Clinical Medicine
基金
国家自然科学基金(81600280)~~
