Figure 4. ROS production in hippocampus and prefrontal cortex.

ROS production was presented as relative of the control group. (a) The hippocampi of chronic unpredictable mild stress exposed mice displayed over-production of ROS when compared to the control group (F_{1, 16} = 42.22, P < 0.0001, post hoc ***P < 0.0001, n = 5, 5, 5, 5), and compared to the hydrogen-rich water group (F_{1, 16} = 10.60, P = 0.0050, post hoc ***P < 0.0001, n = 5, 5, 5, 5). The stress-induced excessive production of ROS in hippocampi could be successfully inhibited by the antioxidative hydrogen-rich water as shown in the comparison between hydrogen-rich water + stress group and stress group (F_{1, 16} = 10.60, P = 0.0050, post hoc ***P < 0.0001, n = 5, 5, 5, 5). (b) Prefrontal cortexes of chronic unpredictable mild stress exposed mice displayed over-production of ROS when compared to the control group (F_{1, 16} = 96.18, P < 0.0001, post hoc ***P < 0.0001, n = 5, 5, 5, 5), and compared to hydrogen-rich water group (F_{1, 16} = 65.54, P < 0.0001, post hoc ***P < 0.0001, n = 5, 5, 5, 5). The stress-induced excessive production of ROS in prefrontal cortexes could be successfully inhibited by the antioxidative hydrogen-rich water as shown in the comparison between hydrogen-rich water + stress group and stress group (F_{1, 16} = 65.54, P < 0.0001, post hoc ***P < 0.0001, n = 5, 5, 5, 5). Representative ROS photographs from hippocampus and prefrontal cortex were shown in (a–d) and (f–i) respectively. No significant differences observed among the control, hydrogen-rich water and hydrogen-rich water + stress groups. Data was analyzed using a two-way ANOVA followed by the Bonferroni’s multiple comparisons post hoc test. Error bars represent s.e.m.