Abstract
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.
Highlights
Polyunsaturated fatty acids (PUFAs) exhibit several favorable effects on cardiovascular diseases [1], autoimmune disorders and inflammatory disorders such as psoriasis [2], rheumatoid arthritis [3,4] and diabetes [5]
For more than 30 years, researchers have known that polyunsaturated fatty acids (PUFAs) can influence the immune system [6]
The results suggested that a low dose of Eicosapentaenoic acid (EPA) up-regulated nitric oxide (NO), inducible nitric oxide synthase (iNOS) and cytokine production via a GPR120-mediated Raf-ERK1/2-IKKβ-NF-κB p65 signaling pathway in RAW264.7 cells
Summary
Polyunsaturated fatty acids (PUFAs) exhibit several favorable effects on cardiovascular diseases [1], autoimmune disorders and inflammatory disorders such as psoriasis [2], rheumatoid arthritis [3,4] and diabetes [5]. For more than 30 years, researchers have known that polyunsaturated fatty acids (PUFAs) can influence the immune system [6]. Itis used as a food supplement with several beneficial effects, including the prevention of cardiovascular diseases, cancers and diabetes [7,8]. EPA and DHA intake as food or supplement effectively prevented cardiovascular events, cardiac deaths and coronary events in patients with high cardiovascular risk [9], it still has some controversy [10,11]. Research has shown that pre-treatment with 100 μmol of EPA decreases LPS-induced pro-inflammatory cytokine production by macrophages [13]. Only a few studies have demonstrated that EPA induces the activation of macrophages
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