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. 2022 Jul 26;13:921553. doi: 10.3389/fphar.2022.921553

FIGURE 5.

FIGURE 5

Hippocampal BDNF-knockdown by BDNF-shRNA abrogated the antidepressant activity of oroxylin A in mice. (A) Fluorescence images of a fixed hippocampal slice which expressed AAV-BDNF-shRNA-EGFP 2 weeks after its stereotactic infusion. The scale bars of representative and enlarged images are 400 and 50 μm, respectively. The following western blotting results confirmed the silencing effects of BDNF-shRNA on the protein expression of hippocampal BDNF (n = 5). (B) Two schematics respectively showing the timeline of the experimental procedures involving CUMS and CRS. (C) Mice in the (CUMS + oroxylin A + BDNF-shRNA)-treated group spent significantly more time being immobile than mice in the (CUMS + oroxylin A)-treated and (CUMS + oroxylin A + Control-shRNA)-treated groups in the FST and TST (n = 12). Also, mice in the (CUMS + oroxylin A + BDNF-shRNA)-treated group displayed notably lower sucrose preference than mice in the (CUMS + oroxylin A)-treated and (CUMS + oroxylin A + Control-shRNA)-treated groups (n = 12). (D) Mice in the (CRS + oroxylin A + BDNF-shRNA)-treated group had significantly more immobility than mice in the (CRS + oroxylin A)-treated and (CRS + oroxylin A + Control-shRNA)-treated groups in the FST and TST (n = 12). Also, mice in the (CRS + oroxylin A + BDNF-shRNA)-treated group exhibited evidently lower sucrose preference than mice in the (CRS + oroxylin A)-treated and (CRS + oroxylin A + Control-shRNA)-treated groups (n = 12). All data were displayed as means ± S.E.M; ** p < 0.01 vs. Vehicle; ## p < 0.01 vs. (CUMS + Vehicle) or (CRS + Vehicle). The statistical comparisons were achieved by (One-way ANOVA + Tukey’s test).