Figure 4.

Propolis-induced wound closure requires hydrogen peroxide. (A) Fluorescence values, corresponding to H₂O₂ increasing concentrations were recorded at 10 min in control cells (ctrl), or in cells incubated with catalase (500 U, CAT, 30 min preincubation), in the presence or not of 0.001% propolis (*** p < 0.001, Bonferroni post-test). Data are presented as means ± SD of rhodamine 123 fluorescence expressed in arbitrary units; n = 16 microplate wells from two different experiments. (B). Fluorescence values recorded at 10 min in scrambled cells or in cells with RNAi for AQP3 (AQP3 RNAi), in the presence or not of 0.001% propolis (*** p < 0.001, Bonferroni post-test). Data are presented as means ± SD of rhodamine 123 fluorescence expressed in arbitrary units; n = 16 microplate wells from two different experiments. (C) Measurements of wound closure in control (ctrl) cells or in cells exposed to CAT 500 U, in the presence or not of 0.001% propolis, expressed as the difference between wound width at 0 and 24 h. Bars represent mean ± SD of two independent experiments, each with n = 25. The mean of the control was set to 100 (*** p < 0.001, Bonferroni post-test). (D) Micrographs of scratch wound. Control (ctrl) cells or cells exposed to CAT 500 U were incubated in the presence of 0.001% propolis and then stained with blue toluidine and observed 24 h after wounding.