Objective:

2

To investigate the effects of acupuncture on the neurotransmitter levels and gut microbiota in a mouse model of Tourette syndrome (TS).

Methods:

2

Thirty-six male C57/BL6 mice were randomly divided into 4 groups using a random number table method: 3

3'-iminodipropionitrile (IDPN) group

control group

acupuncture group

and tiapride group

with 9 mice in each group. In the IDPN group

acupuncture group

and tiapride group

mice received daily intraperitoneal injections of IDPN (300 mg/kg body weight) for 7 consecutive days to induce stereotyped behaviors. Subsequently

in the acupuncture intervention group

standardized acupuncture treatment was administered for 14 consecutive days to IDPN-induced TS model mice. The selected acupoints included Baihui (DU 20)

Yintang (DU 29)

Waiguan (SJ 5)

and Zulinqi (GB 41). In the tiapride group

mice were administered tiapride (50 mg/kg body weight) via oral gavage daily for 14 consecutive days. The control group

IDPN group

and acupuncture group received the same volume of saline orally for 14 consecutive days. Stereotypic behaviors were quantified through behavioral assessments. Neurotransmitter levels

including dopamine (DA)

glutamate (Glu)

and aspartate (ASP) in striatal tissue were measured using enzyme-linked immunosorbent assay. Dopamine transporter (DAT) expression levels were additionally quantified through quantitative polymerase chain reaction (qPCR). Gut microbial composition was analyzed through 16S ribosomal RNA gene sequencing

while metabolic profiling was conducted using liquid chromatography-mass spectrometry (LC-MS).

Results:

2

Acupuncture administration significantly attenuated stereotypic behaviors

concurrently redu

cing striatal levels of DA

Glu and ASP concentrations while upregulating DAT expression compared with untreated TS controls (

P

<

0.05 or

P

<

0.01). Comparative analysis identified significant differences in

Muribaculaceae

(

P

=0.001)

Oscillospiraceae

(

P

=0.049)

Desulfovibrionaceae

(

P

=0.001)

and

Marinifilaceae

(

P

=0.014) following acupuncture intervention. Metabolomic profiling revealed alterations in 7 metabolites and 18 metabolic pathways when compared to the TS mice

which involved various amino acid metabolisms associated with DA

Glu

and ASP.

Conclusions:

2

Acupuncture demonstrates significant modulatory effects on both central neurotransmitter systems and gut microbial ecology

thereby highlighting its dual therapeutic potential for TS management through gut–brain axis regulation.