Interference in Macrophage Balance (M1/M2): The Mechanism of Action Responsible for the Anti-Inflammatory Effect of a Fluorophenyl-Substituted Imidazole

Figure 3

Effect of fluorophenyl-imidazole on TLR-4 receptor (CD284-MD2) (a); mannose receptor (CD206^hi+) (b); phosphorylated protein p65 (c); apoptosis (d); and phagocytic activity (e) in RAW 264.7 macrophages stimulated with LPS (1 μg/mL). The experimental groups used were B: cells pretreated with vehicle and incubated with PBS (blank group); LPS: cells pretreated with vehicle and stimulated with LPS (1 μg/mL); dexa + LPS: cells pretreated with dexamethasone (7 μM) and stimulated with LPS (1 μg/mL); flu (1 μM): cells pretreated with the best concentration (based on IC₅₀ values) (1 µM) of fluorophenyl-imidazole alone; and (flu 1μM) + LPS: fluorophenyl-imidazole (1 µM), before LPS (1 μg/mL). Taxel (30 μM) was used alone in apoptosis experiments, as positive control of apoptosis. The results for TLR-4 (CD284-MD2) and (CD206^hi+) were quantified as receptor expression percentage in relation to cells stained with control isotype antibody while, phosphorylated protein p65 expression was compared to the blank control (cells with no treatment) that represents the basal expression phosphorylated p65 protein. The apoptosis values (%) were quantified as the percentage of apoptotic cells compared with the total cell number and the phagocytic index was measured using the following equation: phagocytic index (PI) = A1/A0; where A1 is the absorbance of sample, LPS and dexamethasone, and A0 is the absorbance of black control. The results were expressed as the mean ± SEM; n = 3; ns, not significant; ; and .

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