Sugmule-4 Decoction for Insomnia: Chemical Composition, Network Pharmacology, and Anti-Insomnia Effects

This study aims to rapidly identify and assign the chemical components in Sugmule-4 decoction using UPLC Orbitrap Exploris MS/MS, conduct network pharmacological analysis, and clarify the anti-insomnia effect of Sugmule-4 decoction. The methods involved using an ACQUITY UPLC BEH C18 (2.1 mm × 100 mm, 1.7 μm) chromatographic column, with 0.1% formic acid aqueous solution (A) - 0.1% formic acid acetonitrile solution (B) as mobile phase gradient elution. The flow rate was 0.5 mL/min, and the injection volume was 5 μL. Under the control of the Xcalibur software (Thermo Fisher Scientific), primary and secondary mass spectrometry data were collected based on the FullScan-ddMS2 function. By combining a self-built database and online databases, the chemical components in Sugmule-4 decoction were identified using accurate relative molecular weight, fragment ion information, pyrolysis pattern, and comparison of control samples. The identified chemical components were then analyzed using network pharmacology. The targets of the drug and insomnia were obtained from TCMSP, SwissTargetPrediction, GeneCards, OMIM, TTD, DrugBank, and DisGeNET databases, respectively, and the intersection target was identified. Protein interaction analysis was performed through the STRING database. The Metascape platform was used to analyze GO function and KEGG pathway, predicting the mechanism of Sugmule-4 decoction in treating insomnia. To determine the anti-insomnia effect of Sugmule-4 decoction, SD rats were divided into a normal control group (K), a model group (M), a Sugmule-4 decoction group (S), and an Anshenbunao liquid group (A). After chronic unpredictable mild stress and PCPA were used for modeling, the rats were given corresponding drugs by gavage for 7 days. The open field text was then conducted to observe the general state and behavioral changes, and the serum levels of AChE, 5-HT, and GABA were measured by ELISA. A total of 136 components were identified from Sugmule-4 decoction, including 37 terpenoids, 34 alkaloids, 26 flavonoids, 20 phenylpropanoids, 14 phenolic acids, and 5 amino acids. Network pharmacology analysis showed that the mechanism of Sugmule-4 decoction in treating insomnia is primarily achieved by its components, such as Romucosine D, Nornantenine, Nuciferine, and (R) - ar Turbone, acting on targets such as ACHE, ADORA2A, and CHRM1 through signal pathways such as Neuroactive ligand-receptor interaction, synaptic signaling, and trans synaptic signaling. This is a multi component-multi target-multi signal pathway joint implementation process. Pharmacodynamic experiments demonstrated that Sugmule-4 decoction and Anshenbunao liquid could revise the general state and behavioral changes of insomnia model rats, increasing the content of AChE, 5-HT, and GABA in serum (P<0.01). This study comprehensively reflects the chemical components of Sugmule-4 decoction and preliminarily summarizes the mass spectrum decomposition characteristics of various chemical components. Network pharmacology and pharmacodynamics experiments show that Sugmule-4 decoction has significant anti-insomnia effects, providing a reference basis for the quality control and mechanism study of Sugmule-4 decoction.