| 摘要: |
| ［摘要］ 目的 探讨黄芩素（BAL）对铁过载引起心肌细胞损伤的保护作用机制。方法 通过200 μmol/L柠檬酸铁（FC）干预心肌细胞H9c2 24 h构建铁过载细胞模型。分析不同浓度BAL（3、10、30 μmol/L）对铁过载的心肌细胞的保护作用，以及抑制Kelch-样ECH相关蛋白1（Keap1）/核因子E2相关因子2（Nrf2）/血红素加氧酶-1（HO-1）通路（经HO-1抑制剂SnPP或Nrf2抑制剂ML385处理）对BAL保护心肌细胞作用的影响。通过CCK-8法检测细胞活力，通过实时定量聚合酶链反应（RT-qPCR）检测炎症因子肿瘤坏死因子α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的表达水平，通过DCFH-DA荧光探针检测活性氧（ROS）水平，采用免疫荧光染色检测4-羟基壬烯醛（4-HNE）表达情况。通过试剂盒检测氧化应激指标丙二醛（MDA）、谷胱甘肽（GSH）表达水平以及铁含量。通过Western blot实验检测Keap1/Nrf2/HO-1通路相关蛋白以及铁死亡相关蛋白表达水平。结果 BAL可提高铁过载H9c2细胞的活力，降低炎症因子TNF-α、IL-1β和IL-6以及氧化应激指标ROS、4-HNE的表达水平，使MDA表达水平降低，GSH表达水平升高，干预效果呈剂量依赖性。经BAL干预后，铁过载H9c2细胞铁沉积减少，铁死亡相关蛋白溶质载体家族7成员11（SLC7A11）、谷胱甘肽过氧化物酶4（GPx4）、溶质载体家族3成员2（SLC3A2）蛋白表达升高，环氧化酶2（COX-2）蛋白表达降低。Western blot实验结果显示，BAL激活Keap1/Nrf2/HO-1通路，抑制Keap1蛋白表达，促进Nrf2、HO-1蛋白表达，干预效果呈剂量依赖性。经SnPP或ML385处理后，BAL对铁过载H9c2细胞的保护作用减弱，细胞活力降低，炎症因子表达升高，氧化应激和脂质过氧化增强，铁沉积增多，铁死亡增强。结论 BAL可能通过激活Keap1/Nrf2/HO-1通路而减轻铁过载引起的心肌细胞损伤。 |
| 关键词: 黄芩素 铁过载 心肌细胞 氧化应激 Keap1/Nrf2/HO-1通路 铁死亡 |
| DOI：10.3969/j.issn.1674-3806.2026.01.14 |
| 分类号:R 285 |
| 基金项目:柳州市科技计划项目（编号：2022SB005） |
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| Baicalein mitigates iron overload-induced cardiomyocyte injury via activating the Keap1/Nrf2/HO-1 pathway |
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LONG Xingjiang, QIN Jiaofeng, HUANG Xiangui.
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Department of Pediatrics, Liuzhou People′s Hospital, Liuzhou 545000, China
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| Abstract: |
| ［Abstract］ Objective To explore the protective mechanism of baicalein(BAL) on cardiomyocyte injury caused by iron overload. Methods An iron-overload cell model was constructed by intervening cardiomyocyte H9c2 with 200 μmol/L ferric citrate(FC) for 24 hours. The protective effects of different concentrations of BAL(3, 10, and 30 μmol/L) on cardiomyocytes with iron overload were analyzed. The protective effects of BAL on cardiomyocytes by inhibiting the Kelch-like ECH-associated protein 1(Keap1)/ nuclear factor erythroid 2-related factor 2(Nrf2)/ heme oxygenase-1(HO-1) pathway(treated with HO-1 inhibitor SnPP or Nrf2 inhibitor ML385) were analyzed. Cell viability was detected by using the CCK-8 assay. The expression levels of inflammatory factors including tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6) were detected by using real-time quantitative polymerase chain reaction(RT-qPCR). The level of reactive oxygen species(ROS) was detected by using 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA) fluorescent probe. The expression of 4-hydroxynonenal(4-HNE) was detected by using immunofluorescence staining. The expression levels of oxidative stress indicators of malondialdehyde(MDA) and glutathione(GSH), and iron content were detected by using reagent kits. The expression levels of proteins related to the Keap1/Nrf2/HO-1 pathway and ferroptosis were detected by using Western blot assay. Results BAL could enhance the viability of iron-overloaded H9c2 cells, reduce the expression levels of inflammatory factors of TNF-α, IL-1β and IL-6, as well as oxidative stress indicators of ROS and 4-HNE, making the expression level of MDA decrease and the expression level of GSH increase, and the intervention effects were dose-dependent. In addition, after intervention with BAL, iron deposition in the iron-overloaded H9c2 cells decreased, while the expressions of ferroptosia-related protein solute carrier family 7 member 11(SLC7A11), glutathione peroxidase 4(GPx4), and solute carrier family 3 member 2(SLC3A2) protein increased, and the expression of cyclooxygenase-2(COX-2) protein decreased. The results of Western blot assay showed that BAL activated the Keap1/Nrf2/HO-1 pathway, inhibited the expression of Keap1 protein, and promoted the expressions of Nrf2 and HO-1 proteins, and the intervention effects were dose-dependent. After treatment with SnPP or ML385, the protective effect of BAL on iron-overloaded H9c2 cells weakened, and the cell viability decreased, and the expressions of inflammatory factors increased, and the oxidative stress and lipid peroxidation enhanced, and the iron deposition increased, and the ferroptosis intensified. Conclusion BAL may mitigate iron overload-induced cardiomyocyte injury via activating the Keap1/Nrf2/HO-1 pathway. |
| Key words: Baicalein(BAL) Iron overload Myocardial cells Oxidative stress Keap1/Nrf2/HO-1 pathway Ferroptosis |
