Pistacia lentiscus Hydrosol: Untargeted Metabolomic Analysis and Anti-Inflammatory Activity Mediated by NF-κB and the Citrate Pathway

Figure 4

Effect of P. lentiscus hydrosol on ATP citrate lyase. (a) U937 cells were transiently transfected with pGL3 basic-LUC vector containing the full-length −3116/−20 bp region of the ACLY gene promoter (3000) or a truncated version of this region (1000). Then, cells were triggered with LPS in the absence (LPS) or in the presence of P.l 1 : 10 or P.l 1 : 100. Unstimulated cells were used as negative control. Twenty-four hours later, the luciferase gene reporter activity was assessed. (b, c) U937 cells, preincubated for 1 hour with P.l 1 : 10 or P.l 1 : 100, were activated with LPS, and ACLY protein levels (b) and enzymatic activity (c) were evaluated. In (b), ACLY and β-actin proteins were immunodecorated with specific antibodies. The Western blotting image (upper panel (b)) is representative of two independent experiments with similar results. The intensities of immunolabeled protein bands were measured by using a quantitative software and normalized to β-actin (lower panel (b)). The bar chart reported of ACLY (b) protein levels obtained from the abovementioned two independent experiments. In (a) and (c), activities are shown as the of three experiments. Statistical significance of differences was evaluated by one-way ANOVA followed by Tukey’s test for multiple comparisons. Different letters indicate significant differences between treatments at .

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