Acanthopanax koreanum Fruit Waste Inhibits Lipopolysaccharide-Induced Production of Nitric Oxide and Prostaglandin E₂ in RAW 264.7 Macrophages

Figure 5

Effect of CH₂Cl₂ fraction of Acanthopanax koreanum fruit waste on the mRNA expression and protein levels of iNOS (a) and COX-2 (b) in LPS-stimulated RAW 264.7 cells. RAW 264.7 cells (5.0 × 10⁵ cell/mL) were stimulated with LPS (1 𝜇g/mL) and the CH₂Cl₂ fraction of A. koreanum (12.5, 25, 50, 100 𝜇g/mL) for 24 hours. The mRNA expression of iNOS and COX-2 was determined by RT-PCR. For the western blot analysis, RAW 264.7 cells (1.0 × 10⁶ cell/mL) were stimulated with LPS (1 𝜇g/mL) in the presence of A. koreanum (12.5, 25, 50, 100 𝜇g/mL) for 24 hours. Whole-cell lysates (25 𝜇g) were prepared and the protein was subjected to 10% SDS-PAGE; expression of iNOS, COX-2 and 𝛽-actin were determined by western blotting. 𝛽-Actin served as a loading control. The mRNA expression of iNOS and COX-2 was determined by RT-PCR.

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