WANG Xuan,LIU Ya-hui,LI Yong,et al.Protective Effect of Daidzein on Senescence of H9c2 Cells Induced by Hydrogen Peroxide[J].Soybean Science,2022,41(04):472-479.[doi:10.11861/j.issn.1000-9841.2022.04.0472]
黄豆苷元对H2O2诱导H9c2细胞衰老的保护作用
- Title:
- Protective Effect of Daidzein on Senescence of H9c2 Cells Induced by Hydrogen Peroxide
- Keywords:
- daidzein; oxidative stress; cell senescence; antioxidant; H2O2; H9c2 cells
- 文献标志码:
- A
- 摘要:
- 为探究黄豆苷元（Daidzein，DAI）对H2O2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（400 μmol?L-1 H2O2）与DAI低、中、高剂量组（20，40，80 μmol?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H2O2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。
- Abstract:
- In order to investigate the protective effect of Daidzein (DAI) on H2O2-induced senescence of rat cardiomyocytes and its mechanism， we randomly divided the H9c2 cells into the control group， model group (400 μmol?L-1 of H2O2)， and low， medium and high-dose DAI groups (20， 40 and 80 μmol?L-1 of DAI， respertively). And we observed the cell senescence by senescence-associated β-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 H2O2. 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.
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备注/Memo
收稿日期：2022-02-14