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. 2022 Jul 6;11(7):1330. doi: 10.3390/antiox11071330

Figure 8.

Figure 8

Effects of thymol, TEOs, and ACHP NFκB inhibitor on mRNA (A,C,E,G) and protein (B,D,F,H) levels of proinflammatory cytokines IL-6, IL-8, IL-1β, and TNF-α after LPS pretreatment. THP-1 cells were pretreated with 100 ng/mL LPS for 24 h and then with 500-fold diluted thymol, TEOs, or 5 μM ACHP for 24 h. DMSO-treated cells were used as control of the treated cells. Real-time PCR for the proinflammatory cytokines was performed with SYBR green protocol. β-actin was used as housekeeping gene, and the relative expression of controls was regarded as 1. Pro-inflammatory cytokine secretions were determined using IL-6-, IL-8-, IL-1β-, and TNF-α-specific ELISA kits according to the manufacturer’s protocols. The columns represent mean values, and error bars represent standard deviation (SD) for three independent experiments (n = 3). The assays were carried out in triplicate in each experiment. Asterisk indicates p < 0.05 compared to control. Cross marks p < 0.05 compared to LPS treatment. Double cross shows p < 0.05 compared to TEO/beginning of flowering. Bullet marks p < 0.05 compared to TEO/end of flowering. Number sign indicates p < 0.05 compared to ACHP treatment. Statistical analysis was carried out by two-way ANOVA followed by Scheffe’s post hoc test.