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

Abstract

Background Ziziphi Spinosae Semen (ZSS) is a plant widely used as medicine and food in Asian countries due to its numerous health benefits. γ-aminobutyric acid (GABA), a non-proteinaceous amino acid, is one of the major inhibitory neurotransmitters with a relaxant function. In this study, a system pharmacology approach was employed to assess the effects of a mixture composed of ZSS and GABA (ZSSG) on sleep improvement. Methods Mice were divided into five groups (n = 10) and received either no treatment, sodium pentobarbital, or sodium barbital with diazepam or ZSSG. The effects of ZSSG on sleep quality were evaluated in mice, and differential metabolites associated with sleep were identified among the control, ZSS, GABA, and ZSSG groups. Additionally, network-based ingredient-insomnia proximity analysis was applied to explore the major ingredients. Results ZSSG significantly improved sleep quality by 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. Furthermore, ZSSG also significantly decreased sleep latency-induced by sodium barbital in mice (P < 0.05). Metabolic analysis revealed significant differences in 10 metabolites between ZSSG group and the groups administering ZSS or GABA. Lastly, using the network-based ingredient screening model, we discovered potential four active ingredients and three pairwise ingredient combinations with synergistic effect on insomnia from ZSSG among 85 ingredients identified by UPLC-Q/TOF–MS. Also, we have constructed an online computation platform. Conclusion Our data demonstrated that ZSSG improved the sleeping quality of mice and helped to balance metabolic disorders-associated with sleep disorders. Moreover, based on the network-based prediction method, the four potential active ingredients in ZSSG 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 or symptoms, such as insomnia.

Network medicine‐based analysis of the hepatoprotective effects of Amomum villosum Lour. on alcoholic liver disease in rats

“…To construct a network involving drug targets and disease‐related proteins, we first mapped the ingredient target proteins and ALD proteins to the human interaction network. Subsequently, we extracted a sub‐network to generate the drug–disease subgraph utilizing the Python NetworkX package, as previously described (Airong Ren et al., 2024 ; Ren et al., 2023 ). The extracted node and edge lists were subsequently imported into Gephi software (version 0.9.2) to generate a comprehensive drug–disease diagram.…”

“…Subsequently, the samples were centrifuged at 4000× g for 20 min. The supernatant was characterized by a Waters Ultra‐High‐Performance Liquid Chromatography‐Quadrupole/Time‐of‐Flight–Tandem Mass Spectrometry (UPLC‐Q/TOF–MS/MS), as previously described (Ren et al., 2024 ; Ren et al., 2023 ). Chromatographic separation was performed on a Waters Acquity™ UPLC system using an ACQUITY UPLC BEH C18 column.…”

Traditional Chinese Medicine in Colorectal Cancer Therapy: A Focus on <i>Panax quinquefolium L.</i> through Text Mining and Network Medicine Approaches

“…When the Z-Score < -1.5 and p ≤ 0.05, the TCM ingredients were considered to have a pharmacologic action. For more details, please refer to our previous research [19,20].…”

Combined Effects of Ziziphus jujuba, Dimocarpus longan, and Lactuca sativa on Sleep-Related Behaviors through GABAergic Signaling

“…By screening through a tobarbital-induced sleep test in a mouse model, it was reported that R. multiflora volved in changes in sleep latency and duration [54]. In addition, in the case of Z. j seeds and walnut peptide, it was reported that the combined use of GABA improves quantity by inducing changes in neurotransmission through pentobarbital-induced test, molecular biological techniques, and ultra-high performance liquid chrom raphy-mass spectrometry analysis [55,56]. In addition, the analysis of sleep latency duration reported differences in sleep-promoting effects depending on the type of le The effects of exposure to Z. jujuba, D. longan, and L. sativa extracts and mixed extracts on changes in sleep-related behavior in fruit flies were validated using ICR-mice, a vertebrate animal (Figure 7).…”

“…By screening through a pentobarbital-induced sleep test in a mouse model, it was reported that R. multiflora is involved in changes in sleep latency and duration [54]. In addition, in the case of Z. jujuba seeds and walnut peptide, it was reported that the combined use of GABA improves sleep quantity by inducing changes in neurotransmission through pentobarbital-induced sleep test, molecular biological techniques, and ultra-high performance liquid chromatographymass spectrometry analysis [55,56]. In addition, the analysis of sleep latency and duration reported differences in sleep-promoting effects depending on the type of lettuce and is also used as a tool to verify expected active ingredients [38,50].…”