【摘要】 目的 构建过氧化氢（H₂O₂）诱导的早衰人脐静脉内皮细胞（HUVEC）模型，观察白藜芦醇能否抑制细胞早衰，并从SIRT1信号通路角度探讨其潜在作用机制。方法 培养HUVEC,CCK-8实验筛选白藜芦醇给药剂量。HUVEC分为对照组、模型组、白藜芦醇低、中、高浓度（10、20、30μmol/L）组，以H₂O₂（0.2μmol/L）诱导建立HUVEC早衰模型，检测细胞存活率、细胞内活性氧（ROS）、丙二醛（MDA）含量、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GPX）活性；SA-β-半乳糖苷酶染色实验观察衰老阳性率；流式细胞术检测细胞凋亡率；实时荧光定量聚合酶链反应（qRT-PCR）和蛋白印迹法（Western blot）检测细胞中沉默信息调节因子1（SIRT1）、检测衰老信号通路中肿瘤蛋白P53（P53）、细胞周期蛋白依赖性激酶抑制因子1A（P21）、细胞周期蛋白依赖性激酶抑制因子2A（P16）、视网膜母细胞瘤抑制蛋白（Rb）mRNA和蛋白表达。采用siRNA干扰技术沉默SIRT1基因表达，观察SIRT1对白藜芦醇抑制H₂O₂诱导HUVEC早衰的影响。结果 与模型组比较，白藜芦醇组细胞存活率显著上升（P<0.05）,衰老细胞阳性率和细胞凋亡率降低（P<0.05）。同时，白藜芦醇干预可使ROS水平和MDA含量显著降低（P<0.05）,SOD和GPX活性显著增加（P<0.05）,SIRT1表达水平升高（P<0.05）,衰老信号通路P53、P21、P16、Rb表达下降（P<0.05）。结论 白藜芦醇能够通过激活SIRT1信号通路抑制H₂O₂诱导HUVEC早衰发生。更多还原

【Abstract】 Objective To establish an H₂O₂-induced senescence model in human umbilical vein endothelial cells（HUVECs）, evaluate whether resveratrol inhibits cellular senescence, and elucidate the involvement of the SIRT1 signaling pathway.Methods HUVECs were cultured and resveratrol dosing was determined by CCK-8 viability assay. Cells were assigned to control, H₂O₂ model, and resveratrol treatment groups（10, 20, 30μmol/L）. Senescence was induced with H₂O₂（0.2μmol/L）. Outcomes included cell viability, intracellular reactive oxygen species（ROS）, malondialdehyde（MDA）, activities of antioxidant enzymes（SOD, GPX）, senescence-associated β-galactosidase（SA-β-gal） positivity, and apoptosis（flow cytometry）. Expression of SIRT1 and senescence-related markers（p53, p21, p16, Rb） was quantified by qRT-PCR and Western blot. To confirm mechanism, SIRT1 expression was silenced by siRNA and the effects of resveratrol were reassessed. Results Resveratrol significantly improved cell viability and reduced SA-β-gal positivity and apoptosis compared with H₂O₂ alone（P<0.05）. It lowered ROS and MDA levels while increasing SOD and GPX activities（P<0.05）. Resveratrol upregulated SIRT1 and downregulated p53, p21, p16, and Rb at both mRNA and protein levels（P<0.05）. Importantly, SIRT1 knockdown by siRNA attenuated the protective effects of resveratrol against H₂O₂-induced senescence, indicating that SIRT1 activation mediates the anti-senescence action of resveratrol. Conclusion Resveratrol inhibits H₂O₂-induced senescence in HUVECs through activation of the SIRT1 signaling pathway.更多还原