恩格letin抑制炎症和铁死亡，并减轻缺氧-复氧诱导的心肌细胞损伤。
Engeletin Inhibits Inflammation and Ferroptosis and Attenuates Cardiomyocyte Injury Induced by Hypoxia-Reoxygenation.
作者信息
Yao Yili, Ling Jianghong, Wang Xiaolong
机构信息
Department of Cardiovascular (Internal Medicine).
Department of Gastroenterology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, 200021, China.
出版信息
Acta Cardiol Sin. 2025 May;41(3):361-371. doi: 10.6515/ACS.202505_41(3).20250221A.
BACKGROUND
Investigate the function and mechanism of engeletin in myocardial ischemia reperfusion injury (MIRI).
METHODS
Hypoxia-reoxygenation (HR) was achieved by subjecting H9c2 cells to 2 hours of hypoxia followed by 4 hours of reoxygenation. The viability of the H9c2 cells was measured by cell counting kit-8 assay. The expressions of interleukin-1 beta (IL-1β), interleukin-6 (IL)-6 and tumor necrosis factor-alpha (TNF-α) were detected by reverse transcription polymerase chain reaction. Reactive oxygen species (ROS) generation was detected by cell-permeable fluorogenic probe Dichloro-dihydro-fluorescein diacetate. Malondialdehyde, superoxide dismutase (SOD) and glutathione (GSH) levels were measured by corresponding kits. The accumulation of intracellular iron ions was accurately measured by the Iron Assay kit. Cell apoptosis was detected by Annexin V-FITC/Propidium Iodide staining. Protein expression was detected by Western blotting to investigate the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear factor-kappa B (NF-κB) signaling pathways.
RESULTS
Engeletin treatment reversed the cell viability induced by HR, and also alleviated cell inflammation by inhibiting the expression of inflammatory cytokines, specifically IL-1β, IL-6 and TNF-α. Furthermore, engeletin treatment significantly inhibited the ROS generation induced by HR, inhibited MDA expression, and promoted SOD and GSH expressions. In addition, engeletin treatment decreased the intracellular concentration of ferrous iron, and promoted both glutathione peroxidase 4 and solute carrier family 7 member 11 expressions. The cell apoptosis results illustrated that engeletin significantly inhibited the apoptosis induced by HR. The Western blotting results showed that engeletin could activate the Nrf2 pathway and downregulate the NF-κB pathway. Engeletin alleviated MIRI in a left anterior descending artery mouse myocardial infarction model.
CONCLUSIONS
Engeletin functioned as a dual regulator both on NF-κB and Nrf2 pathways to alleviate the cell inflammation and ferroptosis induced by HR.
背景
研究恩格letin在心肌缺血再灌注损伤（MIRI）中的作用及机制。
方法
通过对H9c2细胞进行2小时缺氧及随后4小时复氧来实现缺氧复氧（HR）。采用细胞计数试剂盒-8法检测H9c2细胞的活力。通过逆转录聚合酶链反应检测白细胞介素-1β（IL-1β）、白细胞介素-6（IL）-6和肿瘤坏死因子-α（TNF-α）的表达。用细胞可渗透的荧光探针二氯二氢荧光素二乙酸酯检测活性氧（ROS）的产生。用相应试剂盒检测丙二醛、超氧化物歧化酶（SOD）和谷胱甘肽（GSH）水平。用铁测定试剂盒精确测量细胞内铁离子的积累。通过膜联蛋白V-异硫氰酸荧光素/碘化丙啶染色检测细胞凋亡。通过蛋白质印迹法检测蛋白质表达，以研究核因子红细胞2相关因子2（Nrf2）和核因子κB（NF-κB）信号通路的激活情况。
结果
恩格letin处理可逆转HR诱导的细胞活力，并通过抑制炎性细胞因子特别是IL-1β、IL-6和TNF-α的表达减轻细胞炎症。此外，恩格letin处理显著抑制HR诱导的ROS产生，抑制丙二醛表达，并促进SOD和GSH表达。此外，恩格letin处理降低了细胞内亚铁离子浓度，并促进了谷胱甘肽过氧化物酶4和溶质载体家族7成员11的表达。细胞凋亡结果表明，恩格letin显著抑制HR诱导的凋亡。蛋白质印迹结果显示，恩格letin可激活Nrf2通路并下调NF-κB通路。恩格letin在左前降支动脉小鼠心肌梗死模型中减轻了MIRI。
结论
恩格letin作为NF-κB和Nrf2通路的双重调节剂，可减轻HR诱导的细胞炎症和铁死亡。
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