Synergistic Enhancement of Anti-aging Effects on Human Umbilical Vein Endothelial Cells Treated With the Combination of Ferulic Acid and Rapamycin

INTRODUCTION: Aging is a complex process involving cellular, genetic, metabolic, and mitochondrial changes. While significant progress has been made in understanding aging mechanisms and developing anti-aging drugs, single-drug treatments have limitations. This paper aims to investigate the synergistic effects of Ferulic acid (FA) and Rapamycin (Rapa) on anti-aging and to elucidate their underlying mechanisms, providing novel strategies for future anti-aging therapies. METHODS: The safe concentration ranges of FA and Rapa for Human umbilical vein endothelial cells (HUVECs) were determined via Cell counting kit (CCK-8) and Senescence-associated β- Gal staining, with EC50 calculated by GraphPad Prism 8.0.2. Effects on cell cycle arrest and ROS in D-gal-induced aging HUVECs were assessed, with synergistic mechanisms explored by Western Blot and RT-qPCR for aging markers, inflammatory factors, and fibrosis genes. RESULTS: CCK-8 showed that 20-160 μM FA and 50-200 pM Rapa enhanced HUVECs proliferation, with EC50 of 37.78 μM for FA and 48.32 pM for Rapa. The optimal 1:2 combination ratio demonstrated reduced G0/G1 cells, decreased ROS, and lowered NF-κB p65, p53, IL-1β, and TNF-α expression. It also inhibited fibrosis-related gene transcription, downregulating aging markers and maintaining cellular homeostasis. DISCUSSION: These results align with previous studies highlighting FA's antioxidant properties and Rapa's role in mTOR inhibition, suggesting that their combination targets multiple aging pathways simultaneously. The dual approach-reducing oxidative damage while modulating inflammation and fibrosis-may offer superior efficacy compared to single-drug interventions. CONCLUSION: In summary, this dual-target strategy presents a promising avenue for developing advanced anti-aging therapies, warranting further investigation in preclinical and clinical settings.