Exploring the Therapeutic Mechanism of Jianpi Zhidong Decoction on Tourette Syndrome Based on Proteomics and Network Pharmacology

Ning Zhang,1,* Hongxian Zhang,1,* Jianning Guo,2 Yaluan Ma,3 Xue Bai,1 Ning Ma,1 Xiaoxiao Ji,1 Yanli Meng,1 Huifang Li,1 Tananan Sangwanit,1 Yixin Shi,1 Jing Zhao,1 Xiang Li,1 Jingyuan Lin,1 Xia Cui1 1Pediatric Department, Beijing University of Chinese Medicine Third Affiliated Hospital, Beijing, People’s Republic of China; 2Pediatric Department, China-Japan Friendship Hospital, Beijing, People’s Republic of China; 3Laboratory of Molecular Biology, Institute of Basic Theory of Traditional Chinese Medicine, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xia Cui, Department of Pediatrics, Beijing University of Chinese Medicine Third Affiliated Hospital, No. 51 Xiaoguan Street, Andingmen Wai, Chaoyang District, Beijing, 100029, People’s Republic of China, Email [email protected]: To explore the pharmacological effects of Jianpi Zhidong Decoction (JPZDD) on Tourette Syndrome (TS) using proteomics and network pharmacology.Materials and Methods: Chemical components of JPZDD were identified via UPLC-MS/MS. Chronic restraint stress TS model was established by intraperitoneal injection of iminodipropionitrile (IDPN) for 1 week with restraint stress for 3 weeks. Sixty male SD rats were divided into control, model, Tiapride (Tia), and JPZDD groups. After the intervention of 28 days, behavioral tests, Nissl staining, Western blot, immunofluorescence, colorimetry, and ELISA were performed to evaluate the pharmacological effects of JPZDD. Proteomics and network pharmacology predicted targets, validated by Western blot.Results: JPZDD alleviated stereotypic behaviors, hippocampal pathology, and modulated glucose metabolites (GLU, pyruvate, lactate, ATP). It downregulated GLUT1, GLUT3, HK2, and LDHA levels while upregulating PDHA level. Besides, JPZDD balanced M1/M2 microglial phenotypes, reducing IL-1β and IL-6 and increasing IL-4 and IL-10. UPLC-MS/MS identified 44 active ingredients and 123 targets; proteomics revealed 28 differentially expressed proteins. GO/KEGG analysis implicated that the PI3K/AKT/mTOR pathway may be the molecular target. JPZDD inhibited PI3K, AKT, and mTOR phosphorylation.Conclusion: JPZDD (16 g·kg⁻¹·d⁻¹) alleviates motor tics, modulates microglial activation and glucose metabolism, and downregulates the PI3K/AKT/mTOR pathway, providing a mechanistic basis for its therapeutic role in TS. Keywords: Tourette syndrome, Jianpi Zhidong decoction, glucose metabolism, microglia, network pharmacology