Eicosapentaenoic Acid (EPA) Induced Macrophages Activation through GPR120-Mediated Raf-ERK1/2-IKKβ-NF-κB p65 Signaling Pathways.
Eicosapentaenoic Acid (EPA) Induced Macrophages Activation through GPR120-Mediated Raf-ERK1/2-IKKβ-NF-κB p65 Signaling Pathways.
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DOI：
10.3390/nu9090937
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发表时间：
2017-08-25
期刊：
影响因子：
5.9
通讯作者：
Wang C
中科院分区：
文献类型：
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作者：
Han L;Song S;Niu Y;Meng M;Wang C
目的：探讨二十碳五烯酸（EPA）对RAW264.7细胞的免疫调节作用及其分子机制。研究方法：本研究采用细胞培养、细胞增殖实验、形态学观察、一氧化氮（NO）测定、细胞因子测定、核蛋白提取、蛋白质印迹分析、NF-κ B p65免疫荧光分析等方法。结果如下：结果表明，EPA能提高RAW264.7细胞的增殖指数，促进细胞因子和一氧化氮（NO）的释放。Western blotting结果显示，EPA处理后RAW264.7细胞中GPR120蛋白水平明显升高。同时，EPA提高了Raf的磷酸化状态，Raf可能是EPA诱导磷酸化ERK 1/2的上游调节因子。此外，磷酸化ERK 1/2还可促进IKK β在胞浆中的表达，并将NF-κ B p65亚基转位到细胞核中，从而调节诱导型一氧化氮合酶（iNOS）和细胞因子的产生。结论：EPA（0.6-3.0 μ mol）通过GPR120介导的Raf-ERK 1/2-IKK β-NF-κ B p65信号通路激活RAW264.7细胞。
Objectives: To investigate the immunomodulatory effect and molecular mechanisms of Eicosapentaenoic acid (EPA, a typical kind of n-3PUFAs) on RAW264.7 cells. Methods: A variety of research methods, including the RAW264.7 cells culture, cell proliferation assays, morphologic observations, measurements of NO production, cytokine assays, nuclear protein extractions, western blot analyses and NF-κB p65 immunofluorescence assays were used in this study. Results: The results showed that EPA could increase the proliferation index and enhance the release of nitric oxide (NO) and cytokines in RAW264.7 cells. Western blotting results revealed that the protein level of GPR120 increased significantly in RAW264.7 cells after EPA treatment. Meanwhile, EPA elevated the phosphorylation status of Raf, which may act as an upstream regulator of EPA-induced phosphorylated ERK1/2. In addition, the phosphorylated ERK1/2 may then promote IKKβ in endochylema and translocate the NF-κB p65 subunit into the nucleus, thus regulating the production of inducible nitric oxide synthase (iNOS) and cytokines. Conclusions: EPA (0.6–3.0 μmol) activates RAW264.7 cells through GPR120-mediated Raf-ERK1/2-IKKβ-NF-κB p65 signaling pathways.