Anti-Aging Potential of Caffeoyl-Spermidine Derivatives from Lycium ruthenicum Murr.: Insights from UPLC-QTOF-MS/MS, Caenorhabditis elegans, and Mechanistic Studies

Abstract

Lycium ruthenicum 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 Caenorhabditis elegans 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 N1,N10-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 daf-2 downregulation and daf-16 upregulation. Further genetic experiments in daf-2 and daf-16 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 insight into how caffeoylation confers enhanced bioactivity to the spermidine scaffold.