摘要
为探究黄豆苷元(Daidzein,DAI)对H_(2)O_(2)诱导的大鼠心肌细胞衰老的保护作用及其机制,将H9c2细胞随机分成对照组、模型组(400μmol·L^(-1)H_(2)O_(2))与DAI低、中、高剂量组(20,40,80μmol·L^(-1)DAI),采用β-半乳糖苷酶(SA-β-gal)染色法观察H9c2细胞衰老程度,采用比色法检测细胞总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,采用DCFH-DA荧光染色法检测细胞中活性氧(ROS)水平,采用Rh123荧光染色法检测线粒体膜电位变化,采用western blotting法检测H9c2细胞p53、p21、Kelch样环氧氯丙烷相关蛋白-1(Keap1)、核因子E2相关因子(Nrf2)、血红素氧合酶-1(HO-1)蛋白表达水平。结果显示:DAI降低了H_(2)O_(2)诱导的衰老H9c2细胞SA-β-gal阳性率以及p53、p21蛋白表达水平,提高了衰老H9c2细胞T-AOC水平和SOD的活性,降低了细胞中MDA的含量及ROS的水平,维持了线粒体膜电位的稳定,抑制了细胞中Keap1蛋白表达并促进了Nrf2及HO-1蛋白表达。研究表明DAI可延缓细胞衰老,增强细胞抗氧化能力,抑制细胞氧化应激,此作用可能与DAI调节Nrf2/HO-1信号通路有关。
In order to investigate the protective effect of Daidzein(DAI)on H_(2)O_(2)-induced senescence of rat cardiomyocytes and its mechanism,we randomly divided the H9c2 cells into the control group,model group(400μmol·L^(-1)of H_(2)O_(2)),and low,medium and high-dose DAI groups(20,40 and 80μmol·L^(-1)of DAI,respertively).And we observed the cell senescence by senescence-associatedβ-galactosidase(SA-β-gal)staining.The total antioxidant capacity(T-AOC),superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were detected by the colorimetric assay.The ROS level was detected by DCFH-DA fluorescence staining.The mitochondrial membrane potential was detected by the Rh123 fluorescence staining.The expressions of p53,p21,kelch-like epichlorohydrin-associated protein-1(Keap1),nuclear factor E2-associated factor(Nrf2),heme oxygenase-1(HO-1)proteins in H9c2 cells were detected with the western blotting method.The results showed that DAI decreased the positive staining of SA-β-gal,down-regulated the expression of p53 and p21 proteins,increased the T-AOC level and SOD activity,decreased the MDA and ROS levels,and maintained the stability of mitochondrial membrane potential in aging H9c2 cells induced by H_(2)O_(2).DAI inhibited the expression of Keap1 protein,up-regulated Nrf2 and HO-1 protein expression.It is suggested that DAI could delay cellular senescence,enhance cellular antioxidant capacity and inhibit cellular oxidative stress.The mechanism may be related to regulation of Nrf2/HO-1 signaling pathways.
作者
王萱
刘亚辉
李勇
金明
WANG Xuan;LIU Ya-hui;LI Yong;JIN Ming(Medical College,Yanbian University,Yanji 136200,China;The Third Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China;Yanbian University Hospital,Yanji 136200,China)
出处
《大豆科学》
CAS
CSCD
北大核心
2022年第4期472-479,共8页
Soybean Science
基金
国家自然科学基金(82060537)。
