Figures index : Anti-inflammatory Effects of Moutan Cortex Radicis Extract, Paeoniflorin and Oxypaeoniflorin through TLR Signaling Pathway in RAW264.7 Cells : Science and Education Publishing

Figures index

Journal of Food and Nutrition Research. 2018, 6(1), 26-31 doi:10.12691/jfnr-6-1-5

Figure 1. Effect of the MCR extract, paeoniflorin, and oxypaeoniflorin on the cytotoxicity of RAW264.7 cells. Viability of cells treated for 24 h with the MCR extract, paeoniflorin, and oxypaeoniflorin. Absorbance at 570 nm was recorded in an ELISA plate reader. (A) the MCR extract; (B) paeoniflorin; (C) oxypaeoniflorin. Values with different letters are significantly different, p < 0.05. The experiment was performed in hexaplicate

Figure 2. Effect of the MCR extract, paeoniflorin, and oxypaeoniflorin on nitric oxide (NO) production and protein expression of iNOS in LPS-induced RAW264.7 cells. Cells were pre-treated with the MCR extract (50 and 100 μg/ml), paeoniflorin (10 and 30 μM), and oxypaeoniflorin (10 and 30 μM) for 4 h and then co-treated with LPS (1 μg/ml) for 18 h. (A) the production of NO in LPS-induced RAW264.7 cells with the presence or absence of the MCR extract, paeoniflorin and oxypaeoniflorin using the Griess reagent at 540 nm. (B) The expression of iNOS mRNA using RT-PCR. (C) The expression of iNOS protein using western blot. Values with different letters are significantly different, p < 0.05. The experiment was performed in triplicate

Figure 3. The MCR extract, paeoniflorin, and oxypaeoniflorin suppress the production of inflammatory cytokines and their mRNA expression in LPS-induced RAW264.7 cells. Cells were pre-treated with the MCR extract, paeoniflorin, or oxypaeoniflorin for 4 h and then stimulated for 18 h with LPS (1 μg/ml). The production of inflammatory cytokines in supernatants was measured using ELISA kit at 450 nm and 560 nm. (A) The production of IL-1β; (B) The production of IL-6; (C) The production of TNF-α. (D) The mRNA expression of pro-inflammatory cytokines was measured by RT-PCR. Values with different letters are significantly different, p < 0.05. The experiment was performed in tetraplicate and triplicate

Figure 4. The MCR extract, paeoniflorin, and oxypaeoniflorin attenuate the downstream targets of TLR4 signaling pathway in LPS-induced RAW264.7 cells. Cells were pre-treated with the MCR extract, paeoniflorin, or oxypaeoniflorin for 4 h and then stimulated with LPS (1 μg/ml). The mRNA expression of pro-inflammatory cytokines was measured by RT-PCR. The protein level of MAPKs was measured using western blot. The experiment was performed in triplicate

Figure 5. The MCR extract, paeoniflorin, and oxypaeoniflorin attenuate LPS induced the phosphorylation of MAPKs in RAW264.7 cells. Cells were pre-treated with the MCR extract, paeoniflorin, and oxypaeoniflorin for 4 h and then stimulated for 30 min with LPS (1 μg/ml). The protein levels of MAPKs were measured using western blot. The experiment was performed in triplicate