Triple-Marker Screening of 20 Terpenes Identifies Candidate Modulators of Senescence-Associated Phenotypes in D-Galactose-Treated Endothelial Cells

Aging is a major risk factor for chronic diseases, including cardiovascular diseases. Endothelial cells (ECs) are particularly vulnerable to age-related stress, and endothelial senescence contributes to vascular aging. In the present study, we applied a triple-screening strategy to identify terpenes that modulate senescence-associated phenotypes in endothelial cells. Rat aortic endothelial cells (RAECs) were exposed to D-galactose to induce a senescence-associated phenotype, and 20 g/L was selected for subsequent experiments. Subsequently, twenty terpenes were tested at four concentrations (10−9, 10−8, 10−7, and 10−6 mol/L) using three phenotypic readouts: senescence-associated β-galactosidase (SA-β-Gal) activity, cell viability, and oxidative stress. The screening identified multiple terpenes that reduced SA-β-Gal activity and oxidative stress, while five compounds improved cell viability under D-galactose conditions. Among them, citral, terpinolene, and farnesol were the only compounds that showed concordant beneficial effects across the three screening endpoints at the same concentration. In addition, 1R-(-)-myrtenol and trans-caryophyllene showed lower SA-β-Gal activity and preferentially reduced cell viability in D-galactose-treated cells, warranting follow-up studies to determine whether they exhibit senolytic activity. Overall, these findings identify terpene candidates that modulate senescence-associated phenotypes in ECs and support further mechanistic studies to define their senescence-related actions and potential relevance for natural-product-based strategies targeting EC dysfunction.