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巨噬源性泡沫细胞中p62蛋白上调作用和机制的研究

林张军,  李倩,  章越凡,  芮耀诚

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文章导航 > 药学实践与服务  > 2019 >  37(5): 400-405,426
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]. 药学实践与服务, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
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]. 药学实践与服务, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
shu
LIN Zhangjun, LI Qian, ZHANG Yuefan, RUI Yaocheng. A study on the role and mechanism of upregulated p62 protein in macrophage-derived foam cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
Citation: LIN Zhangjun, LI Qian, ZHANG Yuefan, RUI Yaocheng. A study on the role and mechanism of upregulated p62 protein in macrophage-derived foam cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
shu

巨噬源性泡沫细胞中p62蛋白上调作用和机制的研究

doi: 10.3969/j.issn.1006-0111.2019.05.004
  • 1.

    海军军医大学药学院药理学教研室, 上海 200433

  • 2.

    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

基金项目: 国家自然科学基金（30672455）

A study on the role and mechanism of upregulated p62 protein in macrophage-derived foam cells

  • 1.

    Department of Pharmacology, School of Pharmacy, Naval Medical University, Shanghai 200433, China

  • 2.

    Department of Pharmacy, No. 901 Hospital of Joint Logistics Support Force of PLA, Hefei 230031, China

  • 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。
    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.
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  • 收稿日期:  2019-06-23
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巨噬源性泡沫细胞中p62蛋白上调作用和机制的研究

doi: 10.3969/j.issn.1006-0111.2019.05.004
  • 1. 海军军医大学药学院药理学教研室, 上海 200433
  • 2. 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
    • 基金项目:  国家自然科学基金（30672455）
  • 收稿日期: 2019-06-23
  • 修回日期: 2019-07-23

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。

English Abstract

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]. 药学实践与服务, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
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]. 药学实践与服务, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
shu
LIN Zhangjun, LI Qian, ZHANG Yuefan, RUI Yaocheng. A study on the role and mechanism of upregulated p62 protein in macrophage-derived foam cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
Citation: LIN Zhangjun, LI Qian, ZHANG Yuefan, RUI Yaocheng. A study on the role and mechanism of upregulated p62 protein in macrophage-derived foam cells[J]. Journal of Pharmaceutical Practice and Service, 2019, 37(5): 400-405,426. doi: 10.3969/j.issn.1006-0111.2019.05.004
shu

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