Mechanism of An Shen Ding Zhi Fang in an IDPN-Induced Rat Model of Tic Disorders: A Proteomics-Based Investigation

Objective: To evaluate the therapeutic effects of the traditional Chinese medicine (TCM) formula, An Shen Ding Zhi Fang (ASDZF), in a rat model of tic disorders (TD) induced by 3,3'-Iminodipropionitrile (IDPN), and to elucidate the underlying molecular mechanisms by examining its effects on the striatal proteome using quantitative proteomics. Method: Male Sprague-Dawley (SD) rats were used to establish a TD model via intraperitoneal injection of IDPN. Following successful model establishment, the rats were randomly assigned to a model group, a tiapride group, and low-, medium-, and high-dose ASDZF groups. A normal control group was also included. All intervention groups received continuous oral gavage for 28 days. Stereotyped behaviors were assessed. The serum levels of dopamine (DA), 5-hydroxytryptamine (5-HT), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). Pathological changes in the striatum were observed using hematoxylin and eosin (H&E) staining. Tandem mass tag (TMT)-based quantitative proteomics, combined with parallel reaction monitoring (PRM) for targeted validation, was applied to analyze the effects of ASDZF on the striatal proteome of TD rats. Results: Compared with the model group, ASDZF significantly ameliorated stereotyped behaviors (P<0.05), reduced the elevated serum levels of DA, 5-HT, and TNF-α(P<0.05), and mitigated the IDPN-induced pathological damage in the striatum, including neuronal disorganization, pyknosis, edema, and glial proliferation. TMT analysis identified 179 differentially expressed proteins (DEPs) between the medium-dose ASDZF group and the model group. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that these DEPs were significantly enriched in pathways related to the dopaminergic synapse and glutamatergic synapse. ASDZF significantly modulated the expression of key proteins associated with vesicular transport, including Rab3a, Rab3gap2, and Arf3, which was subsequently validated by PRM. Conclusion: ASDZF exerts therapeutic effects on the IDPN-induced TD rat model. Its mechanism of action is multi-targeted and involves multiple pathways, including the regulation of neurotransmitter balance, inhibition of neuroinflammatory responses, and protection of striatal neurons.