Effect of PGSP on inflammatory mediator production in RAW264.7 macrophages.

(A) NO concentration in the medium was measured using Griess reagent. (B–D) PGE2, COX-2, IL-1β, IL-6, and TNF-α production was determined using an ELISA kit. All values are presented as the mean ± SD of three independent experiments (n = 3). A different letter (p < 0.05) reveals statistically significant differences within treatments.</p

Effect of PGSP on cytokine expression in RAW264.7 macrophages.

The mRNA expression of cytokines was determined by real-time qPCR. All values are presented as the mean ± SD of three independent experiments (n = 3). A different letter (p < 0.05) reveals statistically significant differences within treatments.</p

Effect of PGSP on the viability of RAW264.7 macrophages.

Cell viability was determined by the EZ-Cytox Cell Viability Assay Kit. All values are presented as the mean ± SD of three independent experiments (n = 3). A different letter (p < 0.05) reveals statistically significant differences within treatments.</p

Effect of PGSP on cytokine expression in RAW264.7 macrophages.

The mRNA expression of iNOS (A) and COX-2 (B) was determined by real-time qPCR. (C) The protein expression of iNOS and COX-2 was determined by western blotting. All values are presented as the mean ± SD of three independent experiments (n = 3). A different letter (p < 0.05) reveals statistically significant differences within treatments.</p

Effects of PGSP on NF-κB and MAPK activation in RAW264.7 macrophages.

Phosphorylation levels of ERK, JNK, and p38 MAPKs were determined by western blotting. All values are presented as the mean ± SD of three independent experiments (n = 3). A different letter (p < 0.05) reveals statistically significant differences within treatments.</p

Supplementary materials and methods.

The immune-enhancing activity of the combination of Platycodon grandiflorum and Salvia plebeian extracts (PGSP) was evaluated through macrophage activation using RAW264.7 cells. PGSP (250–1000 μg/mL) showed a higher release of NO in a dose-dependent manner. The results showed that PGSP could significantly stimulate the production of PGE2, COX-2, TNF-α, IL-1β, and IL-6 in RAW264.7 cells and promote iNOS, COX-2, TNF-α, IL-1β, IL-4, and IL-6 mRNA expression. Western blot analysis demonstrated that the protein expression of iNOS and COX-2 and the phosphorylation of ERK, JNK, p38, and NF-κB p65 were greatly increased in PGSP-treated cells. PGSP also promoted the phagocytic activity of RAW264.7 cells. All these results indicated that PGSP might activate macrophages through MAPK and NF-κB signaling pathways. Taken together, PGSP may be considered to have immune-enhancing activity and might be used as a potential immune-enhancing agent.</div

Primer sequences used in real-time qPCR.

The immune-enhancing activity of the combination of Platycodon grandiflorum and Salvia plebeian extracts (PGSP) was evaluated through macrophage activation using RAW264.7 cells. PGSP (250–1000 μg/mL) showed a higher release of NO in a dose-dependent manner. The results showed that PGSP could significantly stimulate the production of PGE2, COX-2, TNF-α, IL-1β, and IL-6 in RAW264.7 cells and promote iNOS, COX-2, TNF-α, IL-1β, IL-4, and IL-6 mRNA expression. Western blot analysis demonstrated that the protein expression of iNOS and COX-2 and the phosphorylation of ERK, JNK, p38, and NF-κB p65 were greatly increased in PGSP-treated cells. PGSP also promoted the phagocytic activity of RAW264.7 cells. All these results indicated that PGSP might activate macrophages through MAPK and NF-κB signaling pathways. Taken together, PGSP may be considered to have immune-enhancing activity and might be used as a potential immune-enhancing agent.</div

Effect of PGSP on the phagocytic ability of macrophages.

The cellular uptake of FITC-dextran was determined by flow cytometry. (A) Flow data for the uptake by FITC-dextran of macrophages from one of three independent experiments. (B) The proportion of phagocytic macrophage activity. All values are presented as the mean ± SD of three independent experiments (n = 3). A different letter (p < 0.05) reveals statistically significant differences within treatments.</p

Original western blot gel image data.

The immune-enhancing activity of the combination of Platycodon grandiflorum and Salvia plebeian extracts (PGSP) was evaluated through macrophage activation using RAW264.7 cells. PGSP (250–1000 μg/mL) showed a higher release of NO in a dose-dependent manner. The results showed that PGSP could significantly stimulate the production of PGE2, COX-2, TNF-α, IL-1β, and IL-6 in RAW264.7 cells and promote iNOS, COX-2, TNF-α, IL-1β, IL-4, and IL-6 mRNA expression. Western blot analysis demonstrated that the protein expression of iNOS and COX-2 and the phosphorylation of ERK, JNK, p38, and NF-κB p65 were greatly increased in PGSP-treated cells. PGSP also promoted the phagocytic activity of RAW264.7 cells. All these results indicated that PGSP might activate macrophages through MAPK and NF-κB signaling pathways. Taken together, PGSP may be considered to have immune-enhancing activity and might be used as a potential immune-enhancing agent.</div