Identification of A Novel Antioxidant Tripeptide Ameliorating Skin Photoaging

Background Skin photodamage induced by ultraviolet (UV) exposure is the primary factor of photoaging, and developing bioactive molecules targeting this process represents a key focus in dermatology and cosmetic biotechnology. However, a comprehensive screening strategy combining combinatorial library construction with multi‐target validation has not yet been fully established. Objective To establish a systematic screening strategy for bioactive peptide molecules and to assess the antioxidant, anti‐photoaging effects of the identified bioactive peptide on skin cells. Methods In this study, 17 out of 20 common amino acids were chosen according to their antioxidant capacity via H 2 O 2 ‐ and UVB‐induced oxidative stress models. Based on these 17 amino acids, a tripeptide library containing 4913 molecules was constructed using a full combinatorial strategy. With skin melanin synthesis (tyrosinase), extracellular matrix degradation (elastase or hyaluronidase), and oxidative stress regulation (Keap1) as core targets, 10 potential bioactive tripeptides were screened out through molecular docking technology. Subsequently, mitochondrial function was examined by detecting mitochondrial membrane potential and mitochondrial superoxide anion levels to evaluate the antioxidant activity of the identified tripeptide. Detection of mRNA levels of antioxidant‐related genes and RNA‐seq analysis were performed to investigate the associated signaling pathways. Additionally, senescence‐associated β‐galactosidase staining was used to assess the anti‐aging capacity. Results In vitro functional validation confirmed that tripeptide FAW exhibited the optimal performance in migration capacity and antioxidant activity for skin cells. Mechanistic studies revealed that FAW may enhance cellular antioxidant capacity by activating the NRF2 / HMOX ‐1 pathway and regulate skin aging‐related signaling pathways. The results showed that FAW could alleviated UV ‐induced skin cell damage and photoaging, and ameliorate passage number‐induced replicative senescence. Conclusion This study established a screening strategy for bioactive peptide molecules, encompassing amino acid screening, tripeptide library construction, multi‐target molecular docking, and functional validation. Notably, tripeptide FAW identified in this study showed promising application potential for skincare.