Fig. 3
Impact of L. nepetifolia leaf essential oil (LNLEO) on the expression levels of MAPK proteins, proinflammatory cytokines, and inflammatory mediators in LPS-elicited RAW 264.7 cells. Murine RAW 264.7 cells (2 × 105 cells/mL) grown in 96-well plates were initially exposed to LPS, 1 μg/mL for 2 h to trigger inflammation and then co-treated with LNLEO at two doses set at 12.5 and 50 μg/mL for 24 h. Bar graphs representing the ratios of phosphorylated to total MAPKs i.e., (A) ERK, (B) JNK, (C) p38 MAPK, and secreted pro-inflammatory cytokine levels like (D) TNF-α, (E) IL-1β, and (F) IL-6 in the macrophage cells, spectrophotometrically measured using specialized ELISA Development kits. The mRNA expression levels of inflammatory mediators such as (G) iNOS and (H) COX-2 were analyzed using RT-qPCR. Results were quantified and normalized by the 2-ΔΔCT method, with β-actin serving as the internal housekeeping reference gene. Each experimental value is shown as mean ± SD from three independent trials (n = 3). The statistical significance of data across the groups was assessed employing One-way ANOVA followed by Tukey’s post hoc multiple comparison tests. Significance levels are indicated as #p < 0.001 for comparison between the untreated and LPS-treated group, and *p < 0.05, **p < 0.01, ***p < 0.001 for comparisons between the LPS-treated and LNLEO-treated group.
