The role of kinins in the proliferation of fibroblast primed with TNF in scratch wound assay: Kinins and cell proliferation.

Abstract The aim of this study was to evaluate the involvement of both B 1 and B 2 kinins receptors (B 1 R and B 2 R) in the fibroblast proliferation induced by the cytokine tumour necrosis factor (TNF) attempting to establish an in vitro model of wound healing. Murine fibroblasts L-929 were cultivated in 24 wells plaque until total confluence (DMEM (Vitrocell®); 5% fetal bovine serum, 5% CO 2 , 37 °C) and then submitted to the scratch assay. The cells were treated with PBS, TNF (2 ng/mL) and/or mr-TNF antibody (200 μg/mL), or PDTC. The cells received the second set of treatment (3 h later): PBS; 1 μM HOE-140; 1 μM des-Arg 9 -Leu 8 -BK (DALBK) or 100 μM PDTC. TNF was able to increase the cell proliferation when compared with the group treated with PBS. The co-treatment with the TNF antibody completely reversed the TNF effect. The TNF-proliferative effect was blocked by B 1 (DALBK) and B 2 (HOE-140) kinin receptor antagonists administered separately or along, suggesting the involvement of both receptors in the TNF mechanism of action. Furthermore, the treatment with a NF-ĸB inhibitor PDTC completely blocked the cell proliferation. The TNF cell proliferation was incremented with BK (1 μM) treatment, and its effect was totally reversed by HOE-140 treatment. No effect was observed for TNF plus DABK. Eventually, TNF treatment was able to increase TNF level in the growing medium; however, this increase was suppressed by BK treatment. These results suggest that TNF induces cell proliferation and the induced signalling cascade has the B 2 R participation. All these events seem to be totally dependent on the NF-ĸB activation. These inflammatory mediators can improve the wound healing in the resolution of inflammation.