Oxidative Stress Response in HUVECs

HUVECs (Sigma-Aldrich; Merck KGaA) were maintained in endothelial growth medium (Sigma-Aldrich; Merck KGaA) at 37°C with 5% CO₂. HUVECs were pretreated with Mtp (0.1, 1, 10, 100 or 1,000 µM; dissolved in deionized water; purity >98%; Chengdu Herbpurify Co., Ltd.) for 24 h at 37°C. H₂O₂ has been widely used to induce an oxidative environment in vascular endothelial cell models in vitro (16 (link)). To induce cell senescence, HUVECs were treated with 100 µM H₂O₂ for 12 h at 37°C. HUVECs were incubated with 100 ng/ml phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich; Merck KGaA) for 12 h at 37°C to activate NF-κB, as previously described (17 (link)).