Integrating animal experiment, mass spectrometry and network-based approach to reveal the improving sleep quality of Ziziphi Spinosae Semen and γ-aminobutyric acid mixture

Abstract Background Ziziphi Spinosae Semen (ZSS), which is widely consumed as a medicine and food dual-use plant in some regions of Asian countries, has various health benefits. γ-aminobutyric acid (GABA), a non-proteinaceous amino acid, is one of the major neurotransmitter inhibitors that has a relaxant function. In this study, a system pharmacology approach was proposed to evaluate the effects of a mixture composed by ZSS and GABA (ZSSG) on sleep improvement. Methods Mice were divided into 5 groups (n = 10) and received either no treatment or sodium pentobarbital or sodium barbital with Diaz or ZSSG. The effects of ZSSG on improving sleep quality were evaluated in mice. The major ingredients of ZSSG were analyzed by UPLC-Q/TOF-MS. Network-based ingredient-insomnia proximity was applied to explore its major ingredients. Results The results showed that ZSSG displayed significant effects on decreasing sleep latency and prolonging sleep duration in sodium pentobarbital-induced sleeping mouse model (P < 0.05). ZSSG significantly enhanced the brain content of GABA in mice. In addition, ZSSG also significantly decreased sleep latency-induced by sodium barbital in mice (P < 0.05). With the network-based ingredient screening model, four active ingredients and three pairwise ingredient combinations were discovered from ZSSG among 85 ingredients in ZSSG identified by UPLC-Q/TOF-MS. Conclusion These data demonstrated that ZSSG improved the sleeping quality of mice. The potential four active ingredients and three pairwise ingredient combinations could serve as quality markers associated with insomnia. The network-based framework may open up a new avenue for the discovery of active ingredients of herbal medicine for treating complex chronic diseases, such as insomnia.