Improvement of the Fermentation Liquid of Rose roxburghii and Gastrodia elata Fruit Preserves on Insomnia in Mice

In this experiment, Rose roxburghii and Gastrodia elata fermentation liquid (RGFL) was used as the main material to investigate the effects of RGFL on DL-4-chlorophenylalanine (PCPA)-induced sleep deprivation in mice. A mouse model of insomnia was established by intraperitoneal injection of 450 mg/kg PCPA suspension for three consecutive days. Several assessments were conducted, including the pentobarbital sodium-induced sleep test, open field test, and the measurement of serum levels of superoxide dismutase (SOD), malondialdehyde (MDA), interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). Additionally, changes in the brain levels of glutamic acid (Glu), gamma-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT), and dopamine (DA) were analyzed to evaluate the effects of RGFL. Finally, histopathological examination of whole-brain tissue was performed to assess the reversal of brain damage by RGFL. The results showed that, after inducing sleep in mice using a suprathreshold dose (45 mg/kg) of sodium pentobarbital, the model group mice had a sleep onset rate of only 70%, a prolonged sleep latency (6.33±1.0 min) and a shortened sleep duration (23.20±17.13 min), and that the RGFL intervention significantly (P<0.05) increased the sleep onset rate (100%), shortened the sleep latency (3.29±1.98 min) and prolonged the sleep duration (82.0±19.64 min) of the insomnia-prone mice in the PCPA group. The open field test revealed that RGFL reduced hyperactivity and manic-like behaviors in insomnia mice. Furthermore, RGFL significantly (P<0.01) decreased the levels of IL-6, IL-1β, TNF-α, and MDA by 34.87%, 24.13%, 25.72%, and 45.74%, and significantly (P<0.01) increased SOD enzyme activity by 50.18%, and it was able to significantly (P<0.01) down-regulate the content of neurotransmitter Glu (39.62%) and up-regulate the content of GABA (66.84%,), 5-HT (63.37%), DA content (88.06%). Histopathological analysis demonstrated that RGFL alleviated and improved pathological features in the hippocampal region, such as neuronal disorganization, edema, nuclear pyknosis, and vacuolization, partially reversing brain damage. In conclusion, RGFL effectively improved sleep deprivation induced by PCPA in mice, showing superior efficacy compared to the single administration of Rose roxburghii fermentation liquid（RFL） or Gastrodia elata fermentation liquid (GFL) alone. This indicates a synergistic effect.