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Silibinin Suppresses Mediators of Inflammation through the Inhibition of TLR4-TAK1 Pathway in LPS-induced RAW264.7 Cells

Ji-Hyeon Song, Kui-Jin Kim, Boo-Yong Lee

Journal of Food and Nutrition Research. 2016, 4(8), 515-521 doi:10.12691/jfnr-4-8-5
  • Figure 1. Effect of silibinin on the cell viability in RAW264.7 cells. (A) Chemical structure of silibinin. (B) Cell viability was determined using the MTT assay as described in the Materials and Methods
  • Figure 2. Silibinin suppresses the NO production through the regulation of iNOS, COX2, and IL-1β proteins. (A) The production of NO in LPS-induced RAW264.7 cells. (B) Western blot analysis of the iNOS in LPS-induced RAW264.7 treated with 0, 15, 30, and 60 κM of silibinin. (C) Time dependent analysis of iNOS protein in LPS-induced RAW 264.7 cells. (D) Western blot analysis of the pro-inflammatory cytokines including COX2 and IL-1β in LPS-induced RAW264.7 cells treated with silibinin
  • Figure 3. Silibinn decreases TLR4-TAK1expression response to LPS in RAW264.7 cells. (A) TLR4 was proved for antibody against to TLR4 and α-tubulin as a loading control. (B) Western blot were performed by incubating total lysates were with p-TAK1, TAK1, (C) p-IRF3, and α-tubulin specific antibodies
  • Figure 4. Silibinin inhibits the expression of MAPKs express in LPS-induced RAW264.7 cells. (A) RAW264.7 cells were pre-treated with indicated concentration of silibinin for 4 hours, and then co-treat with LPS for 30 min. Western blot were conducted by incubating total lysate were with p-p38MAPK, p38MAPK, p-ERK, ERK, p-JNK, and JNK specific antibodies. (B) p-ERK protein was proved for antibodies against to p-ERK and ERK as a loading control
  • Figure 5. Silibinin regulated inflammatory stimuli-induced oxidative stress-associated proteins and inflammation-medicated low pH values in RAW264.7 cells. (A) Western blot were performed by incubating total lysates were with NOX4, G6PDH, CuZnSOD, GR, catalase, and α-tubulin specific antibodies. (B) Evaluation of pH values in LPS-induced RAW264.7 cells