Network pharmacology-based cocktail of four Ginkgo biloba compounds enhances yeast mitochondrial function and longevity

BACKGROUND: Cellular aging is characterized by oxidative stress and mitochondrial dysfunction. Pharmacological inhibition of cellular aging holds significant promise. Ginkgo biloba extract (GBE), a crude extract rich in bioactive natural products, has shown potential in mitigating age-related pathologies pharmacologically, However, its anti-aging effects, active components, and underlying mechanisms remain unclear. METHODS: Using Saccharomyces cerevisiae (BY4741/BY4742), a well-established model for studying cellular aging, we assessed GBE's effects on chronological lifespan, stress resistance, and reactive oxygen species (ROS). Active compounds in GBE aqueous solution were identified via LC-MS/MS and network pharmacology. A four-compound cocktail (G4C: quercetin, rutin, ginkgolide B, and isorhamnetin) was formulated based on pathway enrichment analysis. Mitochondrial function was evaluated via RNA-seq, oxygen consumption rate (OCR), membrane potential (ΔΨm), and mitochondrial ROS levels. RESULTS: GBE extended chronological lifespan in BY4741 by up to 73 %, enhanced oxidative and thermal stress resistance, and reduced ROS by 66 % in BY4741 and by 44 % in BY4742 strains. LC/MS analysis combining network pharmacology identified G4C components targeting longevity-related pathways. While individual compounds lacked efficacy, addition of G4C into medium improved oxidative stress resistance synergistically. G4C also extended lifespan by 40 % and reduced ROS by 46 % in yeast. RNA-seq revealed G4C downregulated oxidative phosphorylation genes. Functionally, G4C reduced mitochondrial ROS, preserved ΔΨm during aging, boosted OCR (basal/maximal respiration, ATP production), and modulated mitochondrial calcium, indicating enhanced mitochondrial function. CONCLUSION: GBE's anti-aging effects stem from synergistic actions of quercetin, rutin, ginkgolide B, and isorhamnetin. This multi-compound cocktail enhances mitochondrial function, offering a novel strategy for combating cellular aging. Our findings support network pharmacology-guided design of multi-component anti-aging therapeutics.