Guipi Tang Combined with Probiotics Ameliorates Sleep Deprivation-Induced Hippocampal Injury and Pentobarbital-Induced Sleep Alterations in Mice

Sleep disturbance is a prevalent health problem associated with gut microbiota dysbiosis, chronic inflammation, and neuronal dysfunction. Here, we show that Guipi Tang (GPT) combined with probiotics is associated with improvements in pentobarbital-induced sleep responses and hippocampal injury in mice. Compared with individual treatments, the combined intervention showed improvements in several measured outcomes and was associated with changes in gut microbial composition, inflammatory markers, and hippocampal injury. 16S rRNA (16S ribosomal RNA) sequencing (n = 5 per group) revealed alterations in gut microbial composition following sleep deprivation, while the combined treatment modulated the relative abundance of several bacterial taxa. Furthermore, serum levels of the pro-inflammatory cytokines TNF-α (tumor necrosis factor-alpha) and IL-1β (interleukin-1 beta) differed significantly among groups (n = 5 per group; F (4,20) = 3.959, p = 0.0159 and F (4,20) = 10.26, p = 0.0001, respectively). Western blot analysis (n = 4 per group) revealed significant differences in the expression of autophagy-related markers, including ATG5 (F (4,15) = 8.319, p = 0.0032), Beclin-1 (F (4,15) = 13.62, p < 0.0001), the LC3-II (F (4,15) = 13.56, p < 0.0001), and p62 (F (4,15) = 5.870, p = 0.0048). Compared with the Model group, combined treatment significantly increased ATG5, LC3-II and Beclin-1 expression, while reducing p62 expression. In addition, the Bcl-2/Bax ratio increased significantly following combined treatment (n = 4 per group, p < 0.01 vs. Model), accompanied by a reduction in TUNEL-positive cells. Changes in Akt phosphorylation were also observed, although the relationship between Akt signaling and autophagy requires further investigation. Collectively, these findings suggest that GPT combined with probiotics is associated with improvements in pentobarbital-induced sleep responses and hippocampal injury in sleep-deprived mice, accompanied by changes in gut microbial composition, inflammatory markers, and Akt-associated autophagy-related signaling.