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The CH2Cl2 Extract Fraction from Ficus erecta var. sieboldii (Miq.) King Suppresses Lipopolysaccharide-mediated Inflammatory Responses in Raw264.7 Cells

Young-Min Ham, Weon-Jong Yoon, Eun Hwa Sohn, Dae Won Park, Hyelin Jeon, Yong-Hwan Jung, Sung Ryul Lee, Se Chan Kang

Journal of Food and Nutrition Research. 2018, 6(6), 356-364 doi:10.12691/jfnr-6-6-2
  • Figure 1. Extraction, fractionation, and isolation from leaves of Ficus erecta. var. sieboldii (Miq.) King
  • Figure 2. Inhibitory effects of CFE administration on protein expression of iNOS and COX-2. Raw264.7 cells (1.0×106 cells/mL) were pre-incubated for 2 h with F. erecta CH2Cl2 fractions (CFE; 0, 25, 50, and 100 µg/mL). Cells were then stimulated with LPS (1 μg/mL) for 24 h. LPS-induced changes in the protein levels of iNOS and COX-2 were determined by Western blot analysis. β-actin was used as a loading control. Data were expressed as a ratio relative to the LPS alone group. *P<0.05 and **P<0.01 (test fraction vs. the LPS alone group). LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2
  • Figure 3. Inhibitory effects of CFE administration on mRNA expression of iNOS and COX-2. Raw264.7 cells (1.0×106 cells/mL) were pre-incubated for 2 h with F. erecta CH2Cl2 fractions (CFE; 0, 5, 10 and 20 µg/mL). Cells were then stimulated with LPS (1 μg/mL) for 24 h. The LPS-induced changes in the mRNA levels of iNOS and COX-2 were determined by PCR. β-actin was used as a loading control. Data are mean ± standard deviation (SD). The value of the LPS alone group was set at 1, and results were expressed as a ratio relative to the LPS alone group. *P<0.05 and **P<0.01 (test fraction vs. the LPS alone group). LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2
  • Figure 4. Inhibitory effects of CFE administration on production of PGE2, TNF-α, IL-1β, and IL-6 in Raw264.7 cells. Raw264.7 cells (1.0×106 cells/mL) were pre-incubated for 2 h with F. erecta CH2Cl2 fractions (CFE; 0, 25, 50, and 100 µg/mL). Cells were stimulated with LPS (1 μg/mL) for 24 h. LPS-induced production/release of PGE2 (A), TNF-α (B), IL-1β (C), and IL-6 (D) were determined by ELISA. Data are mean ± standard deviation (SD). The value of the LPS alone group was set at 1, and results were expressed as a ratio relative to the LPS alone group. *P<0.05 and **P<0.01 (test fraction vs. the LPS alone group). LPS, lipopolysaccharide; PGE2, prostaglandin E2; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; IL-6, interleukin-6
  • Figure 5. Inhibitory effects of CFE administration on mRNA expression of TNF-α, IL-1β, and IL-6 in Raw264.7 cells. Raw264.7 cells (1.0×106 cells/mL) were pre-incubated for 2 h with F. erecta CH2Cl2 fractions (CFE; 0, 5, 10, and 20 µg/mL). Cells were then stimulated with LPS (1 μg/mL) for 24 h. The LPS-mediated changes in mRNA levels of TNF-α, IL-1β, and IL-6 were determined by PCR. Data are mean ± standard deviation (SD). The value of the LPS alone group was set at 1, and results were expressed as a ratio relative to the LPS alone group. *P<0.05 and **P<0.01 (test fraction vs. the LPS alone group). LPS, lipopolysaccharide; TNF-α, tumor necrosis factor- α; IL-1β, interleukin-1β; IL-6, interleukin-6
  • Figure 6. Outline of the isolation scheme for the F. erecta CH2Cl2 fraction. C1 and C2 were identified as syringaresinol (molecular weight = 418.44) and 6, 7-furano-5-methoxy hydrocoumaric acid (molecular weight = 236.22), respectively