Pachymic acid targets PDIA6 to delay mesenchymal stem cell senescence through the G6PD/STAT3 signaling axis and prevent senile osteoporosis.

Stem cell senescence and consequent decline in regenerative potential are drivers of aging and age-related disorders; however, pharmaceutical interventions targeting the delay of stem cell senescence to alleviate aging and age-related disorders remain poorly defined. Poria cocos is a well-known traditional Chinese medicine reputed for its health-promoting and lifespan-extending properties, yet the mechanism of its effects on stem cell senescence and bone aging remains unclear. To identify an anti-aging compound in P. cocos, establish the mechanistic basis of its action through the stem cell senescence theory and strategy, and provide foundation for developing related pharmaceuticals. We established a model of oxidative stress-induced senescence of mesenchymal stem cells (MSCs) to screen anti-aging agents. Pachymic acid (PA) was identified as a promising candidate by the MSC senescence model. Chemical proteomics and co-immunoprecipitation techniques were used to clarify the molecular targets and mechanisms of PA. PA significantly reversed cell cycle arrest, reduced the accumulation of reactive oxygen species, and inhibited the production of senescence-associated β-galactosidase, and expression of the senescence markers p21 and p16, in senescent human MSCs. PA preserved the expression of the pluripotency factors Oct4, Nanog, and Sox2, restored the differentiation potential, and enhanced colony formation. Mechanistically, PA was bound specifically to the D221 and T166 residues of PDIA6, leading to the upregulation of G6PD and STAT3. PA reduced the serum levels of advanced glycation end products and malondialdehyde while increased testosterone levels; promoted bone formation while inhibiting bone resorption, preventing bone loss, and ameliorating bone microstructure deterioration in senile osteoporosis (SOP) mice. PA reversed senescence-related phenotypes of bone marrow MSCs in SOP mice by activating G6PD/STAT3 signaling. These findings elucidate that PA specifically targets PDIA6, alleviating PDIA6-mediated inhibition of G6PD and activating the G6PD/STAT3 signaling axis to counteract hMSC senescence. PA ameliorates SOP by delaying BMSC senescence through this conserved mechanism.