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

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.

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.

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.

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.