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. result: CCK-8 assay showed that 20-160 μM FA and 50-200 pM Rapa enhanced HUVECs proliferation, with EC50 values of 37.78 μM for FA and 48.32 pM for Rapa. The optimal ratio was 1:2. The combination reduced G0/G1 cells, decreased ROS, and lowered NF-κB p65, p53, IL-1β, TNF-α expression. It also inhibited fibrosis-related gene transcription, reducing ROS, downregulating aging markers, and maintaining cellular homeostasis. 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.