Therapeutic Effects of the Radix Ginseng and Semen Ziziphi Spinosae Drug Combination on PCPA-induced Insomnia in Mice

Background and Objective: Sleep is essential for human physiological functions; however, the prevalence of sleep disorders has increased in recent years. Traditional Chinese Medicine has gained widespread attention owing to the reduced observed side effects and significant efficacy following administration. Thus, this study aimed to provide a new treatment option for sleep disorders using a drug pairing of Radix Ginseng and Semen Ziziphi Spinosae Drug pair (R–S). Materials and Methods: A total of 60 mice were randomly divided into six groups. An enzyme-linked immunosorbent assay (ELISA) was used to measure the SS (somatostatin), SP (substance P), np-y (neuropeptide Y), 5-HTR4 (5-hydroxytryptamine (serotonin) receptor 4) and 5-HTR3 (5-hydroxytryptamine (serotonin) receptor 3) levels in the small intestine tissues of the mice. Real-time quantitative PCR (RT-qPCR) was employed to assess the Piezo1 and Piezo2 mRNA expression levels. Hematoxylin and eosin (H&E) staining was used to observe cell morphology, and immunohistochemistry was employed to detect brain-derived neurotrophic factor (BDNF) and glial fibrillary acidic protein (GFAP) expression in the small intestine. Data were analyzed using SPSS 27.0 software. A p-value < 0.05 was considered statistically significant. Results: The treatment groups presented higher levels of np-y (p < 0.01) and lower levels of SS, SP, np-y, 5-HTR4 and 5-HTR3 (p < 0.01) compared to the model group. Meanwhile, the treatment groups showed a decreasing trend in Piezo1 and Piezo 2 mRNA expression, an elevated positive expression of BDNF, and a reduced positive expression of GFAP proteins (p < 0.05, p < 0.01). Conclusion: R–S treatment can regulate 5-HTR/Piezo/BDNF/GFAP to improve the sleep of mice with PCPA (para-chlorophenylalanine)-induced insomnia mice, which provides experimental evidence for studying R–S in the treatment of insomnia.