Analysis of the composition of Anshen Dingzhi Pill and its mechanism on improving cognitive impairment in sleep deprivation model rats.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Anshen Dingzhi Pill (ADP) is a classic prescription in traditional Chinese medicine (TCM) for treating psychiatric disorders, which has definite clinical therapeutic effect on insomnia. However, the pharmacological material basis and potential mechanism of ADP in improving sleep have not yet been studied. AIM OF THE STUDY: The aim of this study is to analyze the chemical composition of ADP and explore its underlying mechanism in improving sleep deprivation (SD)-associated cognitive impairment (CI). MATERIALS AND METHODS: Chemical profiling of ADP was performed using UPLC-Q-Exactive Orbitrap MS/MS, and therapeutic targets and pathways were predicted through network pharmacology. The SD model rat was established in a multi-platform aquatic environment. Behavioral tests, histological examinations, and ELISA were used to evaluate ADP's neuroprotective effects on SD-associated CI. The P2X7/NLRP3 signaling pathway was further verified through in vivo and in vitro experiments. RESULTS: In this study, 46 bioactive ingredients of ADP were identified. By network pharmacology, 175 overlapping targets between ADP and SD-associated CI were screened and P2X7/NLRP3 signaling pathway was enriched. ADP significantly ameliorated behavioral deficits, neuronal integrity, oxidative stress, neuroinflammation, and modulated apoptosis-related proteins of SD model rats. In vivo and in vitro, western blotting confirmed that ADP alleviated SD-associated neuroinflammation by inhibiting the P2X7/NLRP3 pathway. CONCLUSION: For the first time, this study systematically identified the chemical components of ADP and demonstrated that ADP ameliorates SD-associated CI primarily via inhibiting the P2X7/NLRP3 signaling pathway, providing a scientific basis for its clinical application in treating sleep-related disorders.