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To investigate the effect and underlying mechanism of action of Yueju volatile oil (YJVO) in the treatment of high altitude sleep disturbance (HASD) mice, gas chromatography-mass spectrometry (GC-MS) was used to identify the components of YJVO, while network pharmacology was applied to predict the mechanism of action. KM mice were selected and randomly assigned to several groups: the control group (control), the model group (model), and the YJVO treatment groups at low (YJVO-L, 200 μL·m-3), medium (YJVO-M, 400 μL·m-3), and high (YJVO-H, 800 μL·m-3) doses, as well as the diazepam group (DZP, 2 mg·kg-1). Except for the control group, all mice were subjected to a hypobaric oxygen chamber to establish the HASD model. Animal experiments were approved by the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (ethics No.: TEMPOR20230088). Co-sleeping and weight-bearing swimming experiments were conducted to assess the sleep-regulating effects of YJVO. Hematoxylin and eosin (H&E) staining was employed to observe damage in hypothalamic and hippocampal brain tissues. Enzyme-linked immunosorbent assay (ELISA) was utilized to measure levels of melatonin (MT), γ-aminobutyric acid (GABA), interleukin-6 (IL-6), interleukin-1beta (IL-1β), 5-hydroxytryptamine (5-HT), orexin-A, and tumor necrosis factor-α (TNF-α) in both plasma and brain tissues. Concurrently, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were detected to evaluate oxidative stress. Western blot was used to determine the expression of key proteins in the NF-κB/NLRP3 pathway, and immunohistochemistry (IHC) was used to detect the expression of 5-hydroxytryptamine 1A receptor (5-HT1A) in hippocampal and hypothalamic tissues. The GC-MS results revealed that YJVO was identified to contain 68 components. Network pharmacology results indicated that the mechanism of YJVO in treating HASD involves multiple signaling pathways such as AGE-RAGE, TNF, serotonin, and NF-κB. Behavioral experiments indicated that YJVO significantly prolonged the sleep duration, reduced the sleep latency, and bolstered the physical endurance and anti-fatigue capabilities of mice (P < 0.01). H&E staining results showed significant improvement in the pathological damage of the hippocampus and hypothalamus tissues in HASD mice. ELISA results indicated that YJVO increased the concentrations of sleep-inducing neurotransmitters MT and GABA within brain tissues, decreased the levels of wakefulness-inducing neurotransmitters orexin-A and 5-HT in plasma, and attenuated the secretion of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α (P < 0.01). Biochemical results indicated that YJVO could inhibit the production of MDA in the brain tissue of HASD mice and enhance the activity of SOD and GSH-PX (P < 0.01). Western blot results showed that YJVO downregulated the protein expression of phosphorylate nuclear factor-kappa B (p-NF-κB p65)/nuclear factor-kappa B (NF-κB p65), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase 1 (Caspase-1), and IL-1β in the brain tissue of HASD mice (P < 0.01). IHC results demonstrated that YJVO downregulated the protein expression of 5-HT1A in the hippocampus and hypothalamus tissues of HASD mice (P < 0.01). The study findings indicate that YJVO demonstrates potent therapeutic effects in HASD mice, potentially through the modulation of neurotransmitters and the attenuation of neuroinflammation.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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