Isoorientin reduces inflammatory responses in LPS-induced RAW 264.7 macrophages through MAPK/FoxO signaling pathways
In order to investigate the effect and mechanism of isoorientin (ISO) on the inflammatory response of RAW 264.7 cells induced by lipopolysaccharide (LPS), RAW264.7 cells in the logarithmic growth phase were randomly divided into a control group, a model group (LPS 1 μg·mL-1), and an isoorientin administration groups (ISO 2.5-40 μg·mL-1 + LPS). After 24 h of cell culture in each group, the cytotoxicity of ISO in RAW 264.7 cells was evaluated by MTT method, the content of nitric oxide (NO) in cell culture medium was detected by Griess method, the supernatant content of tumor necrosis factor-alpha (TNF-α) was detected by ELISA, the mRNA expres-sion levels of COX-2 was detected by qRT-PCR, and the protein expressions of COX-2, JNK, p-JNK, p38, p-p38, FoxO1, and FoxO3 were detected by Western blotting. The results showed that, compared with the LPS model group, ISO significantly inhibited LPS-induced production of TNF-α (P<0.001) and NO(P<0.01), and no cytotoxicity was found. In addition, the expression levels of COX-2 mRNA(P<0.001) and protein (P<0.05) were significantly inhib-ited dose-dependently by ISO in LPS-stimulated RAW 264.7 cells. The Western blotting results showed that ISO significantly attenuated LPS-induced phosphorylation of JNK/p38 MAPK signaling pathway, and reduced the nucle-ar translocation of FoxO1/3. In conclusion, ISO can inhibit LPS-induced inflammatory reaction in RAW 264.7 cells, and the mechanism may be related to its regulatory role in MAPK/FoxO signaling pathway.
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