Anti-aging potential of caffeoyl-spermidine derivatives from <i>Lycium ruthenicum</i> Murr.: insights from UPLC-QTOF-MS/MS, <i>Caenorhabditis elegans</i> , and mechanistic studies

<i>Lycium ruthenicum</i> Murr. (LRM) is rich in spermidine derivatives (SPDs) that exhibit diverse bioactivities. This study systematically characterized the structural profiles and anti-aging mechanisms of LRM-derived caffeoyl-SPDs using UPLC-QTOF-MS/MS, semi-preparative HPLC purification, and <i>Caenorhabditis elegans</i> models. LRM extracts, predominantly containing non-glycosylated caffeoyl-SPDs and anthocyanins, significantly delayed aging in worms by reducing lipofuscin accumulation, scavenging reactive oxygen species (ROS), and enhancing antioxidant enzyme activities. These effects were validated using a caffeoyl-SPD-enriched fraction purified by semi-preparative HPLC and the standard compound <i>N</i>1, <i>N</i>10-dicaffeoyl spermidine, confirming that caffeoyl-SPDs are key anti-aging and antioxidant constituents. Network pharmacology predicted six key pathways, with qRT-PCR validating insulin/IGF-1 signaling (IIS) pathway modulation through <i>daf-2</i> downregulation and <i>daf-16</i> upregulation. Further genetic experiments in <i>daf-2</i> and <i>daf-16</i> mutants revealed a dual mechanism: a DAF-16-independent capacity to lower ROS, coupled with a strict DAF-16 dependence for inducing antioxidant enzymes. This work establishes LRM-derived caffeoyl-SPDs as promising anti-aging agents and provides mechanistic insights into how caffeoylation confers enhanced bioactivity to the spermidine scaffold.