真核起始因子4E对小鼠动脉粥样硬化发生、发展的炎症调节作用

Inflammatory regulation of eukaryotic initiation factor 4E in the process of mice atherosclerosis

目的探讨真核起始因子4E(eIF4E)在小鼠动脉粥样硬化发生、发展过程中的炎症调节作用。方法分别用200 ng/μl脂多糖(LPS)和20 ng/μl白细胞介素-4(IL-4)诱导RAW264.7细胞12 h后提取mRNA,24 h后提取蛋白,采用实时荧光定量聚合酶链反应(qRT-PCR)和Western blotting检测eIF4E mRNA和蛋白在M1型和M2型巨噬细胞中的表达变化。复制ApoE^-/-动脉粥样硬化小鼠模型,检测小鼠腹主动脉eIF4E mRNA和蛋白表达变化;组织免疫荧光检测eIF4E在主动脉根部中分别与M1型和M2型巨噬细胞中的共表达。结果M1型巨噬细胞eIF4E mRNA和蛋白表达高于M2型巨噬细胞(P<0.05)。在ApoE^-/-小鼠动脉粥样硬化模型中,ApoE^-/-小鼠腹主动脉eIF4E mRNA和蛋白表达高于C57BL/6J小鼠(P<0.05);eIF4E与CD86的共定位表达高于eIF4E与CD206的共定位表达。结论eIF4E在动脉粥样硬化的发展过程中主要在M1型巨噬细胞中起作用,进而对动脉粥样硬化产生影响。

Objective To investigate the inflammatory regulation of eIF4E during the development of atherosclerosis in mice.Methods Cell experiments:RAW264.7 cells were induced by 200 ng/μl lipopolysaccharide(LPS)and 20 ng/μl interleukin 4(IL-4)respectively.Relative mRNA was extracted 12 hours later and protein was extracted 24 hours later.Real-time quantitative polymerase chain reaction(qRT-PCR)and Western blotting were used to detect the expression levels of eIF4E mRNA and protein in M1 and M2 macrophages.Animal experiments:ApoE^-/-atherosclerosis mouse model was constructed.Real-time quantitative polymerase chain reaction(qRT-PCR)and Western blotting were used to detect the expression level of eIF4E mRNA and protein in abdominal aorta of mice.Tissue immunofluorescence was used to detect the expression of eIF4E in M1 and M2 macrophages in aortic sinus.Results The expressions of eIF4E mRNA and protein in M1 macrophages were higher than those in M2 macrophages(P<0.05).The expressions of eIF4E mRNA and protein in the ApoE^-/-mice abdominal aorta were higher than those in the C57BL/6J control group(P<0.05);the co-localization expressions of eIF4E and CD86 were higher than those of eIF4E and CD206.Conclusion eIF4E plays a major role in M1 macrophages and then affects the development of atherosclerosis.