摘要
目的:探究miR-34b-5p在动脉粥样硬化(AS)泡沫细胞形成及炎症反应中的作用及可能机制。方法:检测34例AS患者和20例健康体检者血清中miR-34b-5p和胰岛素样生长因子结合蛋白1(IGFBP1)mRNA的表达差异。使用氧化低密度脂蛋白(ox-LDL)诱导RAW264.7巨噬细胞构建AS泡沫细胞模型,检测细胞中miR-34b-5p和IGFBP1 mRNA表达水平。双荧光素酶报告基因实验验证miR-34b-5p和IGFBP1的相互作用关系。将miR-34b-5p抑制剂(inhibitor)、抑制剂阴性对照(inhibitor-NC)、IGFBP1 siRNA质粒(si-IGFBP1)和siRNA阴性对照(si-NC)分别或共转染至AS泡沫细胞模型,观察泡沫细胞沉积脂质能力及胞内总胆固醇(TC)、IL-1β、IL-6、TNF-α水平。结果:与健康体检者相比,AS患者血清中miR-34b-5p水平显著升高(P<0.05),IGFBP1 mRNA水平显著降低(P<0.05)。ox-LDL处理的RAW264.7巨噬细胞中miR-34b-5p表达水平显著升高(P<0.05),IGFBP1 mRNA表达水平显著降低(P<0.05)。双荧光素酶报告基因实验显示,IGFBP1是miR-34b-5p的靶基因。与inhibitor-NC组比较,inhibitor组RAW264.7巨噬细胞中IGFBP1蛋白表达水平显著升高(P<0.05)。ox-LDL诱导后,下调miR-34b-5p表达可抑制巨噬细胞脂质沉积能力,降低胞内TC含量及IL-1β、IL-6和TNF-α水平;而干扰IGFBP1基因表达可通过增强巨噬细胞脂质沉积能力,提高胞内TC、IL-1β、IL-6和TNF-α水平,并逆转miR-34b-5p inhibitor对巨噬细胞的干预作用。结论:下调miR-34b-5p表达可抑制AS泡沫细胞的形成,并降低其炎症反应,其机制可能通过靶向上调IGFBP1表达实现。
Objective:To investigate the role of miR-34b-5p in formation of atherosclerosis(AS)-associated foam cells and inflammation,and its possible mechanism.Methods:Expression levels of miR-34b-5p and insulin-like growth factor binding protein-1(IGFBP1)mRNA in serum of 34 AS patients and 20 healthy controls were detected.RAW264.7 macrophages were treated with oxidized low-density lipoprotein(ox-LDL)to construct AS foam cell model,and expression levels of miR-34b-5p and IGFBP1 mRNA in foam cells were detected.Interaction between miR-34b-5p and IGFBP1 was verified by dual luciferase reporter gene assay.miR-34b-5p inhibitor(inhibitor),inhibitor negative control(inhibitor-NC),IGFBP1 siRNA plasmid(si-IGFBP1)and siRNA negative control(si-NC)were separately or co-transfected into RAW264.7 macrophages.Lipid deposition ability of foam cells was observed,and levels of intracellular total cholesterol(TC),IL-1β,IL-6 and TNF-αwere detected.Results:Compared with healthy controls,miR-34b-5p was highly expressed in serum of AS patients(P<0.05),while expression of IGFBP1 mRNA was lower(P<0.05).ox-LDL treatment significantly increased expression of miR-34b-5p(P<0.05),while significantly reduced expression of IGFBP1 mRNA in RAW264.7 macrophages(P<0.05).Dual luciferase reporter assay showed that IGFBP1 was the target gene of miR-34b-5p.Compared with inhibitor-NC group,expression of IGFBP1 protein in RAW264.7 macrophages in inhibitor group was increased significantly(P<0.05).After ox-LDL induction,miR-34b-5p inhibitor inhibited lipid deposition ability of macrophages,and decreased levels of intracellular TC,IL-1β,IL-6 and TNF-α.Interfering expression of IGFBP1 gene could reverse the intervention effect of miR-34b-5p inhibitor on macrophages by enhance lipid deposition ability of macrophages,and increase levels of intracellular TC,IL-1β,IL-6 and TNF-α.Conclusion:Down-regulation of miR-34b-5p expression can inhibit the formation of AS-associated foam cells and inflammatory response,and the mechanism may be realized by targeting up-regulation of IGFBP1 expression.
作者
惠慧
吴光鹏
廖梅
李光智
HUI Hui;WU Guangpeng;LIAO Mei;LI Guangzhi(Department of Cardiology,Hainan Hospital of Traditional Chinese Medicine,Haikou 570203,China;Heart Function Room,Hainan Hospital of Traditional Chinese Medicine,Haikou 570203,China)
出处
《中国免疫学杂志》
北大核心
2025年第4期879-884,共6页
Chinese Journal of Immunology
基金
海南省卫生健康委卫生健康行业科研项目(21A200101)。
