Anxiety Model with Dietary Interventions

The anxiety model was established by the elevated open platform (EOP) with a slight adjustment based on a previous study (14 (link)). Mice were exposed to the clear square Plexiglas board (10 cm × 10 cm) at 1 m, 1 h per day for 30 days (Figure 1; Supplementary Figure 1). The control mice were not exposed to the elevated open platform (EOP) modeling, yet their living conditions and environment were consistent with those of the other experimental groups.
Mice were randomly assigned to one of 10 groups based on their body weight on day one: control, model, buspirone, powder, yogurt, milk, C, C + powder, C + yogurt, and C + milk. Control, model, and buspirone were designed to test the stability of our modeling system. C was short for the WP + CP combination. Since the combination will enter the powder, yogurt, or milk market, we set the groups of products containing C as C + powder, C + yogurt, and C + milk. The base for the product was powder, yogurt, and milk, respectively. All the nutrients were given by intragastric administration. The protocols utilized for administering intragastric (IG) treatment to mice subjected to the EOP model are described herein (Figure 1). Mice arriving at the laboratory were designated as “eligible mice” if they weighed between 21–25 g and covered 3,000–5,000 cm in the open field test (OFT; Supplementary Figure 2). This criterion ensured a consistent and standardized selection process. Locomotor activity, assessed by distance in the open field test, served as a prerequisite for inclusion in the study and minimized potential confounding factors affecting the reliability and validity of outcomes. Behavioral assessments were conducted from day 30 through day 41, after which the mice from each group were humanely sacrificed and sampled a week following the conclusion of the behavioral experiment.