Atividade anti-inflamatória da pterocarpanoquinona LQB 118 em macrófagos murinos

Abstract

LQB 118 is a pterocarpanoquinone, a synthetic hybrid molecule from the union of two bioactive natural molecule groups, pterocarpans and naphthoquinones. LQB 118 biological activity has been reported, including anti-inflammatory properties in allergic lung inflammation model. However, there are no LQB 118 anti-inflammatory properties reports using other inflammation model. Thus, the aim of this study was to evaluate the role of pterocarpanoquinone LQB 118 in murine peritoneal macrophages in the inflammatory process. Briefly, female Swiss mice was elicited with an intraperitoneal (i.p.) injection of thioglycollate (4%). After four days, peritoneal macrophages were obtained and cultured at the concentration of 2 × 105 cells/well. Cells were treated with LQB 118 (5 μM, 1 μM, 0.5 μM and 0.1 μM) in the presence or absence of the inflammatory stimulus, zymosan (0.2 mg / mL) for 24 h. Then, the supernatants were collected for quantification of the cytokines (TNF-α, IL-1β and IL6) by ELISA. Cell viability was determined by the MTT method. For action mechanism analysis of the molecule, the macrophages were cultured at the concentration of 1,5 × 106 cells / well and treated as described above. After 24 h, the cells were used for analysis of the expression of molecules involved in inflammation by flow cytometry. For this, the macrophages were incubated with anti-TLR2, antiCD69 and anti-P-p38 antibodies. As expected, zymosan increased inflammatory cytokines evaluated production. On the other hand, LQB 118 treatment was able to significantly reduce levels of the cytokines TNF-α, IL-1β and IL-6 in the cell culture supernatant. Besides that, treatment with LQB 118 was able to reduce CD69 expression, as well as increased p38 MAPK phosphorylation induced by zimosan. In addition, LQB 118 was able to negatively modulate TLR2 expression in the presence of the inflammatory stimulus. The results obtained are independent of cell death, since none of LQB 118 concentrations interfered with macrophages viability. Therefore, this work demonstrated for the first time the anti-inflammatory effect of LQB 118 on murine peritoneal macrophages.