摘要
为探究紫花苜蓿提取物(ME)与山楂叶提取物(HE)复配物的抗氧化应激活性及作用机制,采用紫外分光光度法对ME、HE、ME和HE质量比为6∶4复配物(ME+HE)中的总黄酮、总三萜皂苷和总多糖质量分数进行了测定,通过自由基清除实验、总还原能力测定和细胞内活性氧(ROS)清除实验,评估了ME、HE、ME+HE的体外抗氧化活性,并运用Western Blot实验对ME+HE的抗氧化应激作用机制进行了探究。结果表明,ME、HE和ME+HE中总黄酮质量分数分别为3.60%±0.10%、14.12%±2.73%和7.13%±1.55%;总三萜皂苷质量分数分别为1.90%±0.15%、4.08%±1.17%和2.85%±0.50%;总多糖质量分数分别为12.38%±4.75%、26.27%±0.73%和18.37%±1.83%。质量浓度为5 g/L的ME+HE具有良好的2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐阳离子(ATBS^(+))自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基、羟基自由基清除能力,清除率分别为95.75%、92.74%、76.00%,总还原能力较ME提升了75.51%。ME+HE能显著清除细胞内过量的ROS,且抑制能力均优于单独使用的ME和HE。油酸诱导后,HepG2细胞内核因子E2相关因子2(Nrf2)和血红素氧合酶1(HO-1)蛋白的表达会下调,不同剂量(0.9375、1.8750、3.7500 g/L)的ME+HE处理后,Nrf2和HO-1蛋白表达的水平均上调,Nrf2蛋白表达分别增加了10.5%、48.9%和53.3%,HO-1蛋白表达分别增加了72.0%、82.0%、85.3%,呈现出剂量依赖性。ME+HE可通过Nrf2/HO-1信号提高抗氧化能力而发挥抗氧化应激作用。
In order to explore the anti-oxidative stress activity and action mechanism of the complex of alfalfa extract(ME)and hawthorn leaf extract(HE),the mass fractions of total flavonoids,total triterpenoid saponins and total polysaccharides in ME,HE and their complex(ME+HE)with a mass ratio of ME to HE of 6∶4 were determined by ultraviolet spectrophotometry.The in vitro antioxidant activities of ME,HE and ME+HE were evaluated by free radical scavenging experiments,total reduction ability determination and intracellular reactive oxygen species(ROS)scavenging experiment.The anti-oxidative stress mechanism of ME+HE was investigated by Western Blot.The results showed that the mass fractions of total flavonoids in ME,HE and ME+HE were 3.60%±0.10%,14.12%±2.73%and 7.13%±1.55%,respectively,those of total triterpenoid saponins in ME,HE and ME+HE were 1.90%±0.15%,4.08%±1.17%and 2.85%±0.50%,respectively,and those of total polysaccharides in ME,HE and ME+HE were 12.38%±4.75%,26.27%±0.73%and 18.37%±1.83%,respectively.ME+HE with a mass concentration of 5 g/L exhibited good diammonium2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate)cation(ATBS+)radical,1,1-diphenyl-2-picrylhydrazyl(DPPH)radical,and hydroxyl radical scavenging ability,with the corresponding scavenging rates of 95.75%,92.74%,and 76.00%,respectively.The total reduction ability was 75.51%higher than that of ME.ME+HE could significantly remove excessive ROS in cells,and the inhibitory ability was better than that of ME and HE alone.The expression of nuclear factor E2-related factor 2(Nrf2)and heme oxygenase 1(HO-1)protein in HepG2 cells was down-regulated after oleic acid induction.After treatment with different doses(0.9375,1.8750,3.7500 g/L)of ME+HE,the expression levels of Nrf2 and HO-1 proteins were up-regulated.The expression of Nrf2 protein increased by 10.5%,48.9%and 53.3%,respectively,and that of HO-1 protein increased by 72.0%,82.0%and 85.3%,respectively,showing a dose-dependence.ME+HE could improve antioxidant capacity through Nrf2/HO-1 signaling and play an anti-oxidative stress role.
作者
杨洋
王瑶瑶
李丹琦
高品
刘学贵
YANG Yang;WANG Yaoyao;LI Danqi;GAO Pinyi;LIU Xuegui(College of Chemical Engineering,Shenyang University of Chemical Technology,Shenyang 110142,Liaoning,China;Institute of Functional Molecules,Shenyang University of Chemical Technology,Shenyang 110142,Liaoning,China;Liaoning Province Key Laboratory of Green Functional Molecular Design and Development,Shenyang University of Chemical Technology,Shenyang 110142,Liaoning,China;National-Local Joint Engineering Laboratory for Development of Boron and Magnesium Resources and Fine Chemical Technology,Shenyang University of Chemical Technology,Shenyang 110142,Liaoning,China)
出处
《精细化工》
北大核心
2026年第2期430-438,共9页
Fine Chemicals
基金
辽宁省科技厅项目(2023JH2/101600021)
辽宁省教育厅高等学校基础研究项目(LJ212410149033)。
