摘要
目的探讨白藜芦醇(RSV)在氧化应激环境中对人脐静脉内皮细胞(HUVECs)衰老、增殖及凋亡的影响,并对相关机制进行探索。方法利用β-半乳糖苷酶染色HUVECs衰老的细胞并记录阳性细胞数,观察过氧化氢(H2O2)单独处理以及联合RSV后,细胞衰老情况的变化;采用长时程动态活细胞成像及分析系统检测H2O2单独处理以及联合RSV后细胞增殖能力的变化并绘制生长曲线;利用流式细胞技术检测H2O2单独及联合RSV对HUVECs细胞周期及细胞凋亡情况的影响;采用Western印迹检测相关蛋白表达情况。结果HUVECs经H2O2处理后,相比于H2O2组,RSV组β-半乳糖苷酶染色阳性细胞比例明显减低,细胞增殖能力增强,二者差异显著(P<0.05);相比于RSV组,H2O2组细胞周期多被阻滞在G1期,且细胞增殖相关蛋白(p-AKT和pERK)表达减少,而衰老相关蛋白p53、p21、p16、p-Rb表达明显增多;而RSV可拮抗H2O2对HUVECs的作用,使HUVECs凋亡细胞数显著减少,Survivin蛋白表达明显增多(P<0.05)。结论RSV可抑制氧化应激环境中HUVECs细胞的衰老及凋亡、促进细胞增殖,对HUVECs细胞具有保护作用,其机制与RSV改变相关蛋白表达有关。
Objective To investigate the effects of resveratrol(RSV)on senescence,proliferation and apoptosis of human umbilical vein endothelial cells(HUVECs)in oxidative stress state,and to explore possible mechanisms.Methods The senescent HUVECs were stained withβ-galactosidase and the number of positive cells were recorded after treated with H2O2 and RSV.Long-term dynamic cell imaging system was used to observe the changes of cell proliferation ability.The effects of H2O2 alone and combined with RSV on cell cycle and apoptosis of HUVECs were detected by flow cytometry.Western blot was used to detect the expression of related proteins.Results After treated with H2O2,the proportion of positive cells in RSV group was significantly lower,and the ability of proliferation was significantly enhanced than that in H2O2 group(P<0.05).Compared with that of RSV group,the cell cycle of H2O2 group was arrested in G1 phase,and the expressions of cell proliferation-related proteins such as p-AKT and pERK were decreased,while the expressions of senescence-associated proteins p53,p21,p16 and p-Rb were significantly increased,and the expression of survivin protein was significantly increased(P<0.05).Conclusions The senescence of HUVECs could be induced by H2O2,and H2O2 could inhibit cell proliferation,and promote cell apoptosis simultaneously.RSV could protect HUVECs by altering the expressions of related proteins in oxidative stress state.
作者
陈昌喆
宋晨曦
李彬
张冬
张睿
窦克非
CHEN Chang-Zhe;SONG Chen-Xi;LI Bin(Beijing Anzhen Hospital,Capital Medical University,Beijing 100029,China)
出处
《中国老年学杂志》
CAS
北大核心
2019年第16期4027-4031,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金(No.81370310)
