Abstract:

This study investigated the protective effects and underlying mechanisms of the enzymatic hydrolysate from Camellia chekiangoleosa Hu seed oil （EHCO） on D-galactose （D-gal）- induced senescence in human normal foreskin fibroblasts （HFF）. A cellular senescence model was established by treating HFFs with 60 mg/mL D-gal. Cells were co-treated with D-gal and EHCO （0.1% or 0.5%） for 24 hours, and cell viability and senescence levels were subsequently assessed. The content of ATP and the activity of SOD were measured using ELISA kits. Mitochondrial function and proliferative capacity were evaluated via fluorescent staining for ROS, JC-1, and EDU. The mRNA expression levels of senescence-associated and inflammation-related genes were quantified by RT-qPCR. Furthermore, the protein expression of apoptosis-related and autophagy-related markers, along with the phosphorylation status of P53 and ERK, were analyzed by Western blotting. In EHCO-treated HFF, the mRNA levels of P16, P21, P53, IL-1β, and IL-6, as well as the phosphorylation levels of P53 and ERK, significantly reduce in a dose-dependent manner （P<0.05）. Concurrently, the pro-apoptotic BAX/BCL-2 protein ratio decreases, while the expression of the autophagy marker protein LC3B elevates. This study demonstrates that EHCO, obtained through lipase-catalyzed hydrolysis of Camellia chekiangoleosa Hu, promotes cell proliferation and protects against D-gal-induced senescence in HFF. Mechanistically, EHCO exerts anti-skin-aging effects by improving fibroblast function via the suppression of the ERK-P53 signaling axis and the activation of cellular autophagy.

Key words: enzymatic hydrolysate of Camellia chekiangoleosa Hu seed oil, fibroblasts, autophagy, senescence