生命早期两歧双歧杆菌BD-1干预可缓解注意缺陷多动障碍雌性大鼠幼年期的多动行为

doi:10.12122/j.issn.1673-4254.2025.04.04

摘要/Abstract

摘要：

目的研究生命早期两歧双歧杆菌BD-1干预对幼年期注意缺陷多动障碍（ADHD）雌性自发性高血压大鼠（SHR）模型的影响及机制。方法对雌性新生Wistar-Kyoto（WKY）大鼠（空白对照组，n=8）和SHR（模型组，n=6）灌胃生理盐水，同时对雌性新生SHR灌胃BD-1（10⁹ CFU/d，实验组，n=6）。3周龄时停止灌胃并采集粪便，7周龄时进行旷场实验并采集粪便，随后处死大鼠。对肠道菌群进行16S rRNA测序，对纹状体多巴胺转运体和酪氨酸羟化酶、前额皮质活化小胶质细胞进行免疫荧光染色，流式细胞术检测肠系膜淋巴结、脾脏和血液中Treg细胞数量。结果 SHR在旷场中的运动距离高于WKY（P<0.0001），BD-1干预降低了SHR的运动距离（P<0.01）。SHR纹状体中多巴胺转运体（P<0.05）和酪氨酸羟化酶（P<0.05）表达低于WKY，BD-1干预增加了SHR酪氨酸羟化酶的表达（P<0.05）。BD-1干预减少了SHR前额皮质活化小胶质细胞数（P<0.01）、增加了SHR脾脏中Treg细胞数（P<0.01）。BD-1干预增加了SHR肠道菌群α多样性、降低厚壁菌门/拟杆菌门比值，促进Muribaculaceae生长、抑制Clostridia_UCG-014生长。结论生命早期两歧双歧杆菌BD-1干预通过调节肠道菌群、调节外周免疫、抑制神经炎症、改善多巴胺能神经系统功能，从而缓解宿主后期多动行为，为ADHD的早期预防和精神益生菌的开发应用提供科学依据。

关键词: 注意缺陷多动障碍, 两歧双歧杆菌BD-1, 微生物-肠-脑轴, 生命早期, 神经炎症

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

杨洋, 王凯, 柳鉴修, 周志谟, 贾雯, 吴思谋, 李金星, 何方, 程如越. 生命早期两歧双歧杆菌BD-1干预可缓解注意缺陷多动障碍雌性大鼠幼年期的多动行为[J]. 南方医科大学学报, 2025, 45(4): 702-710.

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.

图/表 8

图1 BD-1干预对多动行为的影响

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.

图 2 BD-1干预对纹状体中多巴胺转运体和酪氨酸羟化酶表达的影响

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.

图3 BD-1干预对前额皮质活化小胶质细胞和肠系膜淋巴结、脾脏及血液中Treg细胞的影响

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.

图4 BD-1干预对肠道菌群多样性的影响

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.

图5 肠道菌群门水平相对丰度前5位柱状图

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

图6 肠道菌群属水平相对丰度前10位柱状图

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

图7 3周龄肠道菌群属水平相对丰度比较

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.

图8 7周龄肠道菌群属水平相对丰度比较

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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