Integrating pharmacokinetics and network pharmacology to reveal mechanism of Shuangxia decoction in the treatment of insomnia

Shuangxia Decoction is a traditional Chinese medicine classic formula used clinically to treat insomnia, consisting of Pinellia ternata (Banxia) and Prunella vulgaris (Xiakucao). The aim of this study is to explore the pharmacokinetic characteristics of the main core components in Shuangxia Decoction based on Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) analysis, as well as to preliminarily investigate its mechanisms of action in treating insomnia using quantitative targeted network pharmacology. In this research, an LC-MS/MS method was established to analyze four main core components in Shuangxia Decoction: rosmarinic acid, 4-hydroxy-3-methoxyphenyllactic acid (HMLA), danshensu, and liquiritigenin. The pharmacokinetic characteristics of these phenolic compounds were investigated after oral administration of Shuangxia Decoction in rats. Network pharmacology and molecular docking were used to identify the underlying mechanism of Shuangxia Decoction in treating insomnia. The results showed that the analysis of the four components was completed within 6 ​min. The T max for danshensu, HMLA, rosmarinic acid, and liquiritigenin were 0.79 ​± ​0.09, 0.63 ​± ​0.12, 0.51 ​± ​0.21, and 0.38 ​± ​0.19 ​h, respectively; their C max were 110.83 ​± ​10.98, 25.20 ​± ​4.13, 37.57 ​± ​7.70, and 22.27 ​± ​8.75 ​μg/L, respectively; and their T 1/2 were 1.20 ​± ​0.24, 0.79 ​± ​0.26, 4.93 ​± ​1.08, and 2.85 ​± ​0.11 ​h, respectively. Danshensu exhibited the highest peak concentration (C max : 110.83 ​± ​10.98 ​μg/L), while liquiritigenin showed the lowest C max (22.27 ± 8.75 ​μg/L), likely due to hydrolysis by intestinal carboxylesterases. Network pharmacology results indicated that the main active components of Shuangxia Decoction exert their effects primarily through neuro-signaling pathways such as the dopaminergic synapse and glutamatergic synapse. This study is the first to explore the pharmacokinetic characteristics of the four core components in Shuangxia Decoction and to provide preliminary predictions of its mechanisms in treating insomnia, laying a foundation for further exploration of its pharmacological mechanisms.