Statistical analysis

To analyze the laboratory data statistically, a Statistical Program for the Social Science Software version 22 (SPSS Inc., Chicago, IL) was performed. To determine the normality of the testing results for the animals, the Kolmogorov–Smirnov technique was used^54,55. One-way analysis of variance (ANOVA) was conducted for all variables⁵⁶. Values are denoted as the mean ± standard error, and then Tukey's posthoc analysis is performed⁵⁷. The overall significance was recognized at 0.05. Correlations among brain CRF gene expression, serum corticosterone, proinflammatory cytokines, brain oxidative stress markers, and immunoexpression were studied using the Pearson correlation coefficient of pairwise comparison⁵⁸. The correlation heatmap was used to identify these correlations between samples obtained by OriginPro, Version 2023b. (OriginLab Corporation, Northampton, MA, USA). Furthermore, the percentage change = [(mean of the control group − mean of the treated group)/mean of control group] × 100%, the percentage improvement = [(mean of the stressed group − mean of the treated group)/mean of control group] × 100⁵⁹.