Early life Bifidobacterium bifidum BD-1 intervention alleviates hyperactivity of juvenile female rats with attention deficit hyperactivity disorder

doi:10.12122/j.issn.1673-4254.2025.04.04

Abstract

Abstract:

Objective To investigate the effects of early life intervention with Bifidobacterium bifidum BD-1 (B. bifidum BD-1) on hyperactivity in a female mouse model of attention deficit hyperactivity disorder (ADHD) and explore the underlying mechanisms. Methods Eight newborn female Wistar-Kyoto (WKY) rats and 6 spontaneous hypertensive rats (SHRs) were gavaged with saline and another 6 SHRs were gavaged with B. bifidum BD-1 (10⁹ CFU) daily for 3 weeks. Open field test of the rats was conducted at 7 weeks, and fecal samples were collected at weaning (3 weeks) and at 7 weeks for 16S rRNA sequencing. Immunofluorescent staining was used to detect dopamine transporter (DAT) and tyrosine hydroxylase (Th) levels in the striatum and activated microglia in the prefrontal cortex. Treg cells in the mesenteric lymph nodes, spleen and blood were analyzed using flow cytometry. Results The SHRs traveled a significantly greater distance in open fields test than WKY rats, and this behavior was significantly attenuated by B. bifidum BD-1 intervention. The expression of DAT and Th in the striatum was significantly lower in the SHRs than in WKY rats, while B. bifidum BD-1 treatment obviously increased Th levels in the SHRs. B. bifidum BD-1 intervention significantly deceased the number of activated microglia and increased Treg cell counts in the spleen of SHRs. The treatment also enhanced α diversity in gut microbiota of the SHRs and resulted in a decreased Firmicutes/Bacteroidota ratio, more active Muribaculaceae growth, and suppression of Clostridia_UCG-014 proliferation. Conclusion Early life intervention with B. bifidum BD-1 alleviates hyperactivity in female SHRs by modulating the gut microbiota and peripheral immune response, suppressing neuroinflammation and improving dopaminergic system function. These findings provide evidence for early prevention strategies and support the development and application of psychobiotics for ADHD.

Key words: attention deficit hyperactivity disorder, Bifidobacterium bifidum BD-1, microbiota-gut-brain axis, early life, neuroinflammation

Yang YANG, Kai WANG, Jianxiu LIU, Zhimo ZHOU, Wen JIA, Simou WU, Jinxing LI, Fang HE, Ruyue CHENG. Early life Bifidobacterium bifidum BD-1 intervention alleviates hyperactivity of juvenile female rats with attention deficit hyperactivity disorder[J]. Journal of Southern Medical University, 2025, 45(4): 702-710.

Figures/Tables 8

Fig.1 Effects of BD-1 intervention on hyperactivity in SHRs. A: Time moving in the OFT.B: Distance moved in the OFT. C: Representative trace maps of the rats. (n=8 in WKY group, n=6 in SHR group, n=6 in SHR-BD-1 group). **P<0.01, ****P<0.0001.

Fig.2 Effects of BD-1 intervention on expressions of DAT and Th in the striatum of the rats. A: Immunofluorescence staining of the striatum. B: IOD of DAT in the striatum. C: IOD of Th in the striatum. *P<0.05.

Fig.3 Effects of BD-1 intervention on the number of activated microglia in the prefrontal cortex and Treg cells in the mesenteric lymph nodes (MLN), spleen and blood of the rats. A: Immunofluorescence staining and calculation of activated microglia in the prefrontal cortex. B: Number of Treg cells/CD4+ cells (%) in the MLN, spleen and blood (n=8 in WKY group, n=6 in SHR group, n=6 in SHR-BD-1 group). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Fig.4 Effects of BD-1 intervention on gut microbiota diversity in the rats. A: α diversity and principal coordinate analysis map of β diversity of gut microbiota at 3 weeks (n=5). B: α diversity and principal coordinate analysis map of β diversity of the gut microbiota at 7 weeks (n=8 in WKY group, n=6 in SHR group, n=6 in SHR-BD-1 group). **P<0.01, ***P<0.001, ****P<0.0001.

Fig.5 The top 5 components at the phylum level in the gut microbiota of the rats.

Fig.6 The top 10 components at the genus level in the gut microbiota of the rats.

Fig.7 Comparison of the top 10 components at the genus level in the gut microbiota at 3 weeks. *P<0.05, **P<0.01.

Fig.8 Comparison of the top 10 components at the genus level in the gut microbiota at 7 weeks. *P<0.05, **P<0.01, ***P<0.001.

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