Magnolia kobus Extract Inhibits Periodontitis-Inducing Mediators in Porphyromonas gingivalis Lipopolysaccharide-Activated RAW 264.7 Cells

Figure 1. The anti-inflammatory pathway of MKE in RAW 264.7 cells.

Figure 2. Effect of (A) MKE (0, 30, 100, and 300 µg/mL) and (B) magnolin (0, 0.45, 1.5, and 4.5 µg/mL) on RAW 264.7 cell viability. MKE and magnolin were used for 24 h of treatment. Values are represented as means ± standard deviation.

Figure 3. Effect of (A) MKE (30, 100, and 300 µg/mL) and (B) magnolin (0.45, 1.5, and 4.5 µg/mL) on RAW 264.7 cell NO production. MKE and magnolin were used for 24 h with LPS. Nitrite formed in the supernatant was measured using Griess’ reagent. Values represent means ± standard deviation. ## p < 0.01 vs. Vehicle control group; * p < 0.05 and ** p < 0.01 vs. LPS-activated control group.

Figure 4. Effects of MKE (30, 100, and 300 µg/mL) on the production of inflammatory mediators in LPS-stimulated RAW 264.7 cells. MKE and 100 ng/mL of LPS was added except for the vehicle control group and incubated at 37 °C for 24 h. Protein expression levels of TNF-α, IL-1β and mPGES-1 were detected by Western blot. The protein band density was measured by utilizing the ImageJ software. β-actin was used as a control. Values represent means ± standard deviation. # p < 0.05 and ## p < 0.01 vs. Vehicle control group; ** p < 0.01 vs. LPS-activated control group.

Figure 5. Effects of magnolin (0.45, 1.5, and 4.5 µg/mL) on the production of Inflammatory mediators in LPS-stimulated RAW 264.7 cells. Magnolin and 100 ng/mL of LPS were added, but not to the vehicle control group, and incubated at 37 °C for 24 h. Protein expression levels of TNF-α, IL-1β and mPGES-1 were detected by Western blot. The protein band density was measured utilizing ImageJ software. β-actin was used as a control. Values represent means ± standard deviation. ## p < 0.01 vs. Vehicle control group; ** p < 0.01 vs. LPS-activated control group.

Figure 6. Effects of MKE (30, 100, and 300 µg/mL) on the production of MMPs in LPS-stimulated RAW 264.7 cells. MKE and 100 ng/mL of LPS was added except for in the vehicle control group and incubated at 37 °C for 24 h. Protein expression levels of MMP-1, MMP-3, MMP-8, MMP-9 and MMP-13 were detected by Western blot. The protein band density was measured by utilizing ImageJ software. β-actin was used as a control. Values represent means ± standard deviation. ## p < 0.01 vs. Vehicle control group; * p < 0.05 and ** p < 0.01 vs. LPS-activated control group.

Figure 7. Effects of magnolin (0.45, 1.5, and 4.5 µg/mL) on the production of MMPs in LPS-stimulated RAW 264.7 cells. Magnolin and 100 ng/mL of LPS was added except for in vehicle control group and incubated at 37 °C for 24 h. Protein expression levels of MMP-1, MMP-3, MMP-8, MMP-9 and MMP-13 were detected by Western blot. The protein band density was measured by utilizing ImageJ software. β-actin was used as a control. Values represented means ± standard deviation. ## p < 0.01 vs. Vehicle control group; * p < 0.05 and ** p < 0.01 vs. LPS-activated control group.

Figure 8. Effects of MKE (30, 100, and 300 µg/mL) on the production of TLR4 and NF-κB factors in LPS-stimulated RAW 264.7 cells. MKE and 100 ng/mL of LPS was added except for in vehicle control group and incubated at 37 °C for 15 min. Protein expression levels of TLR4, pIκBα, IκBα, pp65 and p65 were detected by Western blot. The protein band density was measured by utilizing ImageJ software. β-actin was used as a control for TLR4 protein expression level. Values represent means ± standard deviation. ## p < 0.01 vs. Vehicle control group; ** p < 0.01 vs. LPS-activated control group.

Figure 9. Effects of magnolin (0.45, 1.5, and 4.5 µg/mL) on the production of TLR4 and NF-κB factors in LPS-stimulated RAW 264.7 cells. Magnolin and 100 ng/mL of LPS was added except for in the vehicle control group and incubated at 37 °C for 15 min. Protein expression levels of TLR4, pIκBα, IκBα, pp65 and p65 were detected by Western blot. The protein band density was measured by utilizing ImageJ software. β-actin was used as a control for TLR4 protein expression level. Values represent means ± standard deviation. ## p < 0.01 vs. Vehicle control group; * p < 0.05 and ** p < 0.01 vs. LPS-activated control group.

Figure 10. Effects of MKE (30, 100, and 300 µg/mL) on the production of pTAK1 and MAPK factors in LPS-stimulated RAW 264.7 cells. MKE and 100 ng/mL of LPS was added except for in the vehicle control group and incubated at 37 °C for 15 min. Protein expression levels of pTAK1, TAK1, pERK, ERK, pJNK, JNK were detected by Western blot. Values represent as means ± standard deviation. ## p < 0.01 vs. Vehicle control group; * p < 0.05 and ** p < 0.01 vs. LPS-activated control group.

Figure 11. Effects of magnolin (0.45, 1.5, and 4.5 µg/mL) on the production of pTAK1 and MAPK factors in LPS-stimulated RAW 264.7 cells. Magnolin and 100 ng/mL of LPS was added except for in the vehicle control group and incubated at 37 °C for 15 min. Protein expression levels of pTAK1, TAK1, pERK, ERK, pJNK, JNK were detected by Western blot. Values represent means ± standard deviation. ## p < 0.01 vs. Vehicle control group; ** p < 0.01 vs. LPS-activated control group.