UPLC-Orbitrap MS/MS Analysis and Network Pharmacology of Sugemule-4 Decoction for Anti-Insomnia Effects

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Abstract

Objective: To rapidly identify and assign the chemical components in Sugemule-4 decoction by using UPLC Orbitrap Exploris MS/MS, and then conduct network pharmacological analysis, and clarify the anti-insomnia effect of Sugmule-4 decoction.

Methods: 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. Flow rate: 0.5 mL/min, injection volume: 5μL. Under the control of the control software (Xcalibur, Thermo Fisher Scientific), the primary and secondary mass spectrometry data are collected based on the FullScan-ddMS2 function. Combined with the self-built database and online database, the chemical components in Sugemule-4 decoction are identified by accurate relative molecular weight, fragment ion information, pyrolysis pattern and comparison of control samples. The identified chemical components were analyzed by network pharmacology, and the targets of drug and insomnia were obtained from TCMSP, SwissTargetPrediction, GeneCards, OMIM, TTD, DrugBank and DisGeNET databases respectively, and the intersection target was obtained; Protein interaction analysis was performed through STRING database. Use Metascape platform to analyze GO function and KEGG pathway, and predict the mechanism of Sugmule-4 decoction in treating insomnia. To determine the anti-insomnia effect of Sugmule-4 decoction, SD rats were divided into normal control group (K), model group (M), Sugmule-4 decoction group (S) and 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, then the open field text was conducted to observe the general state and behavioral changes, and the serum levels of AChE, 5-HT and GABA were measured by ELISA.

Results: 136 components were identified from Sugemule-4 decoction, including 37 terpenoids, 34 alkaloids, 26 flavonoids, 20 phenylpropanoids, 14 phenolic acids and 5 amino acids. The results of network pharmacology analysis show that the mechanism of Sugmule-4 decoction in treating insomnia is mainly achieved by its components such as Romucosine D, Nornantenine, Nuciferine, and (R)-arturbone 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. The results of pharmacodynamic experiment showed that Sugmule-4 decoction and Anshenbunao liquid could revise the general state and behavioral changes of insomnia model rats, increase the content of AChE, 5-HT and GABA in serum(P<0.01).

Conclusion: This study comprehensively reflects the chemical components of Sugmule-4 decoction, and preliminarily summarizes the mass spectrum decomposition characteristics of various chemical components. The network pharmacology and pharmacodynamics experiments show that Sugmule-4 decoction has significant anti-insomnia effect, which provides a reference basis for the quality control and mechanism study of Sugmule-4 decoction.