巨噬源性泡沫细胞中p62蛋白上调作用和机制的研究
投稿时间:2019-06-23  修订日期:2019-07-23  点此下载全文
引用本文:林张军,李倩,章越凡,芮耀诚.巨噬源性泡沫细胞中p62蛋白上调作用和机制的研究[J].药学实践杂志,2019,37(5):400~405,426
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作者单位E-mail
林张军 海军军医大学药学院药理学教研室, 上海 200433  
李倩 中国人民解放军联勤保障部队第九〇一医院药剂科, 安徽 合肥 230031  
章越凡 海军军医大学药学院药理学教研室, 上海 200433  
芮耀诚 海军军医大学药学院药理学教研室, 上海 200433 ruiyc163@163.com 
基金项目:国家自然科学基金(30672455)
中文摘要:目的 探讨巨噬源性泡沫细胞中p62蛋白对脂质代谢相关的自噬以及炎症因子表达的影响,为p62在抗动脉粥样硬化中的应用提供参考。方法 利用Ox-LDL刺激RAW264.7细胞的方法模拟巨噬源性泡沫细胞的形成,通过Western blot及实时荧光定量PCR检测巨噬源性泡沫细胞中p62蛋白和mRNA水平。通过Western blot比较p62 siRNA组和对照组中Ox-LDL诱导的LC3剪切、脂滴相关蛋白Plin2和过氧化物酶体相关蛋白PEX2的蛋白水平,通过实时荧光定量PCR比较TNFα和IL-6 mRNA表达水平。结果 Ox-LDL对RAW264.7细胞中p62蛋白以及mRNA水平均有上调作用。干扰p62的表达之后,LC3-Ⅱ蛋白水平降低,Plin2的蛋白水平并无明显变化,PEX2的蛋白水平升高,TNFα和IL-6 mRNA表达升高。进一步研究发现,干扰Nrf2后能明显抑制Ox-LDL对p62的上调作用。结论 巨噬源性泡沫细胞中Ox-LDL通过Nrf2介导p62蛋白的上调,p62可能具有抗动脉粥样硬化的作用。
中文关键词:动脉粥样硬化  p62  自噬  巨噬细胞  炎症因子
 
A study on the role and mechanism of upregulated p62 protein in macrophage-derived foam cells
Abstract:Objective To investigate the effect of p62 protein on the lipid autophagy and the expression of inflammatory cytokines in macrophage-derived foam cells.Methods RAW264.7 cells were stimulated by oxidized low density lipoprotein to mimic the formation of macrophage-derived foam cells.The levels of p62 protein and mRNA in macrophage-derived foam cells were detected by Western blot and real-time qPCR.The protein levels of LC3,Plin2 and PEX2 were measured by Western blot in Ox-LDL treated Raw264.7 cells to determine the effects of p62 on autophagy in macrophage-derived foam cells.The TNFα and IL-6 mRNA levels were detected by real-time PCR.Results Ox-LDL up-regulated both autophagy levels and p62 protein levels in RAW264.7 cells.After interfering the expression of p62,the level of LC3-Ⅱ protein decreased,but the protein levels of lipid droplet-related protein Plin2 did not change significantly.The peroxisome-related protein PEX2 level increased.The expression of TNFα and IL-6 also increased.Further studies showed that Nrf2 interference significantly inhibited the up-regulation of p62 by Ox-LDL.Conclusion Ox-LDL mediates the upregulation of p62 through Nrf2 in macrophage-derived foam cells,and p62 may play a protective role in atherosclerosis.
keywords:atherosclerosis  p62  autophagy  macrophage  inflammatory cytokines
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