Figure 5.

Hesperidin increases TGF-β1 levels and activates its signaling pathway in cultured astrocytes and in the hippocampus. Cortical astrocytes were treated with hesperidin for 2 h and submitted to In-Cell western analysis for TGF-β1 (A). Activation of TGF-β1 signaling pathway was analyzed by Western blotting assays for phospho-SMADs 2/3 (B) and by evaluation of nuclear translocation of SMADs 2/3 (C–E). Levels of phospho-SMADs 2/3 were also analyzed in the hippocampus of adult mice treated with vehicle or hesperidin (F). Hesperidin slightly increases the levels of astrocytic TGF-β1 and strongly induced phosphorylation and activation of the SMADs 2/3 signaling pathway in vitro and in vivo. *p < 0.050 and **p < 0.010. Student’s t test was used in (A,E,F). Comparisons among multiple groups were analyzed using a one-way ANOVA followed by Newman-Keuls post hoc test in (B). n = 4–5 independent astrocyte cultures for the in vitro experiments and n = 4–6 animals per experimental group for the in vivo experiments.