摘要
目的探讨木蝴蝶总黄酮(TFO)的抗氧化活性及其对人脐静脉内皮细胞(HUVECs)氧化应激和凋亡的影响。方法采用DPPH、ABTS和O^(2-)清除、Fe^(2+)和Cu^(2+)螯合、Fe3+和Cu^(2+)还原法测定木蝴蝶总黄酮的抗氧化水平。采用过氧化氢(H_(2)O_(2))诱导HUVECs损伤模型,测定木蝴蝶总黄酮干预处理后细胞活性和超氧化物歧化酶(SOD)、丙二醛(MDA)、乳酸脱氢酶(LDH)、过氧化氢酶(CAT)和活性氧(ROS)的活性。采用Hoechst 33258、JC-1和Annexin V-FITC/PI染色研究木蝴蝶总黄酮对细胞凋亡的影响。通过蛋白质印迹法检测核因子E2相关因子2(Nrf2)、血红素加氧酶(HO-1)和醌氧化还原酶1(NQO-1)的表达,观察木蝴蝶总黄酮抗氧化应激和凋亡的作用机制。结果木蝴蝶总黄酮具有较强的清除自由基和还原金属离子的能力,而金属螯合能力相对较弱。体外细胞实验显示木蝴蝶总黄酮可明显提高HUVECs的存活率(P<0.01,P<0.001),明显增强SOD和CAT的活性(P<0.05,P<0.001),降低LDH、MDA和ROS的水平(P<0.05,P<0.01,P<0.001),降低H_(2)O_(2)诱导的细胞凋亡(P<0.01,P<0.001),提高Nrf2、HO-1和NQO-1蛋白的表达(P<0.05,P<0.01,P<0.001)。结论木蝴蝶总黄酮具有明显的抗氧化活性,可通过Nrf2/HO-1通路缓解H_(2)O_(2)诱导的氧化应激和细胞凋亡。
Objective To investigate the antioxidant activity of total flavonoids from Oroxylum indicum(TFO)and their effects on oxidative stress and apoptosis in human umbilical vein endothelial cells(HUVECs).Methods Firstly,the antioxidant capacity of TFO was assessed using DPPH,ABTS,and O^(2-)radical scavenging assays,along with Fe^(2+)and Cu^(2+)chelation,and Fe3+and Cu^(2+)reduction assays.Secondly,an oxidative injury model was established in HUVECs using hydrogen peroxide(H_(2)O_(2)).Cell viability and the activities or levels of superoxide dismutase(SOD),malondialdehyde(MDA),lactate dehydrogenase(LDH),catalase(CAT),and reactive oxygen species(ROS)were measured.The effects of TFO on apoptosis were investigated using Hoechst 33258 staining,JC-1 staining,and Annexin V-FITC/PI staining.Finally,to explore the underlying mechanism,the protein expression levels of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and NAD(P)H quinone dehydrogenase 1(NQO-1)were detected by Western Blot.Results TFO exhibited potent free radical scavenging and metal ion reducing abilities,although its metal chelating capacity was relatively weak.In vitro cell experiments demonstrated that TFO significantly enhanced HUVEC viability(P<0.01,P<0.001),markedly increased the activities of SOD and CAT(P<0.05,P<0.001),decreased the levels of LDH,MDA,and ROS(P<0.05,P<0.01,P<0.001),reduced H_(2)O_(2)-induced apoptosis(P<0.01,P<0.001),and up-regulated the expression of Nrf2,HO-1,and NQO-1(P<0.05,P<0.01,P<0.001).Conclusion TFO possesses significant antioxidant activity and can alleviate H_(2)O_(2)-induced oxidative stress and apoptosis in HUVECs,potentially via the Nrf2/HO-1 pathway.
作者
张颖茵
彭倩
许俊雅
陈凯然
周冰旋
谢京玲
李敏晶
陈颖楠
张蕾
ZHANG Yingyin;PENG Qian;XU Junya;CHEN Kairan;ZHOU Bingxuan;XIE Jingling;LI Minjing;CHEN Yingnan;ZHANG Lei(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006 Gunangdong,China;Dept.of Pharmacy,The Second Hospital&Clinical Medical School,Lanzhou University,Lanzhou 730000 Gansu,China)
出处
《中药新药与临床药理》
北大核心
2025年第11期1861-1869,共9页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
广东省基础与应用基础研究基金-省企联合基金面上项目(2023A1515220188)。
