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Randomized Controlled Trial
. 2023 Mar 23:14:1158137.
doi: 10.3389/fimmu.2023.1158137. eCollection 2023.

Psychobiotic Lactobacillus plantarum JYLP-326 relieves anxiety, depression, and insomnia symptoms in test anxious college via modulating the gut microbiota and its metabolism

Ruizhe Zhu  1 , Yilin Fang  1 , Hongyu Li  1 , Ying Liu  2 , Jing Wei  1 , Shuwei Zhang  1 , Liwei Wang  1 , Rui Fan  1 , Lingfang Wang  1 , Shengjie Li  1 , Tingtao Chen  1
Affiliations
Randomized Controlled Trial

Psychobiotic Lactobacillus plantarum JYLP-326 relieves anxiety, depression, and insomnia symptoms in test anxious college via modulating the gut microbiota and its metabolism

Ruizhe Zhu et al. Front Immunol. .

Abstract

Introduction: Test anxiety is a common issue among college students, which can affect their physical and psychological health. However, effective interventions or therapeutic strategies are still lacking. This study aims to evaluate the potential effects of Lactobacillus plantarum JYLP-326 on test anxious college students.

Methods: Sixty anxious students were enrolled and randomly allocated to the placebo group and the probiotic group. Both groups were instructed to take placebo and JYLP-326 products twice per day for three weeks, respectively. Thirty unanxious students with no treatments were assigned to a regular control group. The anxiety, depression, and insomnia questionnaires were used to measure students' mental states at the baseline and the end of this study. 16S rRNA sequencing and untargeted metabolomics were performed to analyze the changes in the gut microbiota and fecal metabolism.

Results: The questionnaire results suggested that JYLP-326 administration could relieve the symptoms of anxiety, depression, and insomnia in test anxious students. The gut microbiomes of the placebo group showed a significantly greater diversity index than the control group (p < 0.05). An increased abundance of Bacteroides and Roseburia at the genus level was observed in the placebo group, and the relative abundance of Prevotella and Bifidobacterium decreased. Whereas, JYLP-326 administration could partly restore the disturbed gut microbiota. Additionally, test anxiety was correlated with disordered fecal metabolomics such as a higher Ethyl sulfate and a lower Cyclohexylamine, which could be reversed after taking JYLP-326. Furthermore, the changed microbiota and fecal metabolites were significantly associated with anxiety-related symptoms.

Conclusion: The results indicate that the intervention of L. plantarum JYLP-326 could be an effective strategy to alleviate anxiety, depression, and insomnia in test anxious college students. The potential mechanism underlying this effect could be related to the regulation of gut microbiota and fecal metabolites.

Keywords: depression and insomnia; gut microbiota; lactobacillus plantarum JYLP-326; test anxiety; untargeted metabolomics.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Study flow chart showing recruitment and group assignment.
Figure 2
Figure 2
Effects of L. plantarum JYLP-326 on the gut microbial diversity. (A) Chao1 and Shannon indices of the gut microbiota. (B) PCoA analysis of gut microbiota. (C) Venn diagram of gut microbial species. HP, the normal control group without any treatmeat; P, the P group taking placebo; PB, the PB group taking probiotic L. plantarum JYLP-326. ∗ p < 0.05.
Figure 3
Figure 3
Effects of JYLP-326 on the gut microbial composition. (A) The relative abundance of gut bacteria at the phyla level. (B) The relative abundance of gut bacteria at genus level. HP, the normal control group without any treatment; P, the P group taking placebo; PB, the PB group taking JYLP-326. ∗ p < 0.05.
Figure 4
Figure 4
Fecal metabolome profiling among groups. (A) PCA plots in ESI+ model. (B) PCA plots in ESI- model. (C) Superclass of the identified fecal metabolites. HP, samples of the normal control group (n=25, green box); P, samples of the P group (n=20, purple circle); PB, samples of the PB group (n=20, pink triangle); QC, the quality control samples (n=7, yellow diamond).
Figure 5
Figure 5
Pathway enrichment analysis of differential metabolites. (A, B) Venn diagram of differential metabolites in ESI+ and ESI- modes. A total of two hundred differential metabolites were shown in the Venn diagrams. (C) KEGG enrichment analysis of these changed metabolites. The scatter diagram shows the enrichment of differential metabolites in various pathways.
Figure 6
Figure 6
Spearman correlation analysis among anxiety-behavior, gut microbiota, and fecal metabolites. Spearman’s rank correlation coefficient among anxiety symptom scores, 13 metabolites in Table 2 and top 6 relative abundance of gut microbiota. P values are depicted in red and blue, where red refers to a negative correlation and blue refers to a positive correlation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
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Grants and funding

This research was funded by grants from National Natural Science Foundation of China (Grant no. 82060638 to TC; 82260389 to SL), the Double thousand plan of Jiangxi Province (High-End Talents Project of Scientific and Technological Innovation to TC), and Jiangxi Provincial Natural Science Foundation (20224BAB206078 to SL; 20212BAB206087 and 20212BCJ23043 to LFW).