Fermented Gastrodia elata Bl. Intervenes gut microbiota and amino acid metabolism in zebrafish to promote 5-HT homeostasis against sleep disturbances.

Sleep disturbances (SD) is a prevalent health issue in modern society. Consequences involve negative impacts on numerous aspects of physical, psychological, and daily life. This study aimed to elucidate the protective effects of Fermented Gastrodia elata Bl. (FGE) against pentylenetetrazole (PTZ)-induced SD and uncover the underlying molecular mechanisms. Our findings revealed that FGE significantly attenuated PTZ-induced insomnia behavior, circadian rhythm disturbances, and compromised gut barrier functions. Mechanistically, gut microbiota and 5-HT within the microbiota-gut-brain axis are the key regulators in short-term SD. Fecal microbiota transplantation (FMT) experiment identified gut microbiota as a key mediator and potential therapeutic target for FGE. The comprehensive analysis of 16S rRNA sequencing and metabolomic analysis showed that amino acid metabolism-related pathways as key factors of FGE intervention. Notably, omics joint analysis demonstrated a strong association between the variations of the intestinal microbiota among different groups and the notable alterations in the brain metabolomic landscape. Meanwhile, the remodeling of intestinal microbial structure and metabolites drove the homeostasis of 5-HT levels in different tissues. Importantly, exogenous keystone bacteria supplementation to sleep-deprived zebrafish restored insomnia responses and amino acid metabolism. Targeted amino acid metabolism further confirmed amino acid metabolism as the central mechanistic pathway through which FGE exerts its protective function. Collectively, Collectively, these findings suggested that FGE showed a significant preventative action on short-term SD by intervening gut microbiota and amino acid metabolism in zebrafish to promote 5-HT homeostasis, which offers perspectives into new preventive strategies of traditional Chinese medicine dietary supplements for transient insomnia.