The researchers began with the most basic question in longevity pharmacology: does the compound actually work? When C. elegans animals were raised on nematode growth media supplemented with Rg3, the treated animals lived significantly longer than untreated controls. The effect was robust under the conditions tested, with an effective concentration identified at 10 micromolar. Developmental exposure was not required for the lifespan benefit, which persisted when animals were exposed only at the adult stage, although the benefit was strongest when treatment began early in life.

The team next ruled out the most common confounders in worm feeding studies. Rg3 did not alter the growth of the OP50 bacteria used as a food source, and the animals showed no avoidance of Rg3-supplemented plates, ruling out a simple feeding aversion. The compound also did not reduce progeny production, indicating that the lifespan extension was not accompanied by reduced fertility. The team also tested whether the effect depended on the worm’s own metabolic state, and found that it did not phenocopy dietary restriction or mitochondrial mutants.

With the basic pharmacology established, the researchers turned to the molecular target. Transcriptomic analysis of Rg3-treated animals identified 753 differentially expressed genes, and pathway enrichment analysis pointed strongly toward phosphoinositide signaling, inositol phosphate metabolism, autophagy, and axon guidance. The team then tested whether the compound’s effect depended on known longevity pathways, and found that it did not require DAF-16/FOXO, dietary restriction, germline signaling, or mitochondrial function. Instead, the effect converged on the autophagy machinery itself: knockdown of bec-1, a core component of the autophagy initiation complex, abolished the lifespan benefit, and the compound did not further extend lifespan in vps-34 or vha-3 mutants affecting lysosomal function.

The researchers then asked whether Rg3 preserved neuronal integrity during aging. They examined the PVD multidendritic sensory neurons, which mediate harsh-touch responses and develop age-related morphological decline. In Rg3-treated animals, the PVD neurons retained their well-organized multidendritic architecture significantly longer than in untreated controls, and the animals responded more robustly to harsh-touch stimulation at late adult stages. The team also examined AWC and ASH olfactory neurons, which showed similar preservation of both morphology and chemotaxis behavior in Rg3-treated animals.

The team then turned to neuronal proteostasis, modeling age-related protein aggregation with polyglutamine and α-synuclein. In animals expressing polyglutamine-YFP in neurons, Rg3 treatment significantly reduced the number of aggregates per animal at late adult stages. In a Parkinson’s disease model expressing human α-synuclein in dopaminergic neurons, Rg3-treated animals retained significantly more dopaminergic neurons than untreated controls at late adult stages, suggesting preservation of neuronal survival in the face of proteotoxic stress.

To probe the mechanism of neuronal protection, the researchers used a neuronal autophagy reporter in which a fluorescent protein is delivered to acidic lysosomal compartments. In Rg3-treated animals, the reporter showed increased delivery to acidic compartments at late adult stages, consistent with enhanced autophagic flux. The team then tested whether the compound’s effect required the lysosomal proton pump VHA-3, and found that knockdown of vha-3 abolished the neuronal protection, indicating that the compound acts through lysosomal acidification.

The researchers then connected the neuronal response to the phosphoinositide pathway. The transcriptomic data identified downregulation of genes encoding FYVE-domain proteins, which bind phosphatidylinositol 3-phosphate, and the team found that Rg3 treatment reduced the accumulation of FYVE-domain reporter puncta in neurons. The team also identified the inositol phosphate pathway as a convergent target, and found that the compound’s effect required the inositol phosphate metabolism genes identified in the transcriptomic screen.

The team then probed the connection to TOR signaling. The transcriptomic data identified downregulation of let-363, the worm TOR ortholog, and the team found that Rg3 treatment reduced TOR activity in neurons. The team then tested whether the compound’s effect required the autophagy transcription factor HLH-30, the worm ortholog of mammalian TFEB, and found that knockdown of hlh-30 abolished the lifespan benefit. The team also found that Rg3 treatment promoted nuclear accumulation of HLH-30 in neurons, consistent with activation of the autophagy transcriptional program.

The work provides a new model for how ginsenoside Rg3 supports healthy aging: the compound acts through phosphoinositide signaling to reduce TOR activity, promote nuclear accumulation of HLH-30/TFEB, and enhance lysosomal acidification and autophagic flux, thereby preserving neuronal morphology, function, and proteostasis during aging. The findings identify the lysosome and autophagy machinery as a central target of ginseng’s longevity pharmacology and provide a new framework for how phosphoinositide signaling supports neuronal maintenance during aging.

Subject of Research: Aging Cell

Article Title: Ginsenoside Rg3 Promotes Healthy Aging Through Phosphoinositide‐Associated Lysosomal Regulation in Caenorhabditis elegans

Article References: Yu, H., Li, S., Chen, Y., Zeng, Y., Li, J., Guo, Y., Zhang, W., Zhang, L., You, L., Chen, K., Wu, J., Gan, D., Liu, J.-K., & Zheng, L. (2026). Ginsenoside Rg3 Promotes Healthy Aging Through Phosphoinositide‐Associated Lysosomal Regulation in Caenorhabditis elegans. Aging Cell, 25(10), Article e70755. https://doi.org/10.1111/acel.70755

Image Credits: AI Generated

DOI: 10.1111/acel.70755

Keywords: ginsenoside Rg3, Caenorhabditis elegans, autophagy, lysosome, TOR signaling, phosphoinositide, healthy aging, neurodegeneration, Ginsenoside, Promotes, Healthy, Aging