Metabolomics and network analysis reveal the mechanism of Salvia miltiorrhiza bunge extract in ameliorating cognitive dysfunction in sleep-deprived rats.

Sleep deprivation (SD) causes learning memory and cognitive impairment. Salvia miltiorrhiza Bunge (Danshen, DS), a medicinal plant in the family Labiatae, has been traditionally used for sleep-related disorders. Previous studies have shown that DS can ameliorate SD-induced cognitive impairment. However, the underlying mechanisms for its pharmacological effects remain unclear. This study aimed to evaluate the protective effects and mechanisms of DS extract against cognitive impairment in SD rats. UPLC-QTOF/MS was used to analyze DS extracts. The SD model was constructed utilizing a modified multi-platform aquatic sleep deprivation procedure that lasted 21 days. The Morris water maze test (MWM), hematoxylin and eosin (H&E) staining, and enzyme-linked immunosorbent assay (ELISA) were used to assess learning and memory ability, hippocampus injury, and serum inflammation, respectively. An integrated strategy of serum metabolomics combined with network analysis was used to explore the potential mechanisms by which DS exerts pharmacological effects. Molecular docking and experiments were used for further validation. UPLC-QTOF-MS/MS identified 32 diterpenoids in DS extract. The results showed that DS (1.35 and 2.70 g/kg) significantly improved spatial learning and memory abilities in SD rats while also reducing hippocampus pathological damage and serum inflammation. Serum metabolomics showed that DS modulated 26 differential metabolites, mainly involved in Glycerophospholipid metabolism, Glycerolipid metabolism, Phosphatidylinositol signaling system, and One carbon pool by folate. Network analysis screened 145 putative targets for DS to alleviate SD-induced cognitive impairment, involved in inflammation regulation and metabolic modulation. Integrated analyses of metabolomics and network analysis indicated that PIK3CA was a key target for DS's regulatory effects, primarily engaged in the regulation of phosphatidylinositol phosphate metabolism. Validation experiments revealed that all eight components of DS extracts had a higher binding ability with PIK3CA, and DS restored the SD-induced abnormal expression of PIK3CA. Our study provides new insights into the development of DS as a dietary supplement for treating SD-induced cognitive impairment.