Wang ZiLin, Liu SiMiao, Shi JiaHao, Chen Di, Li ShaoLin, Yu ShengQin, Liu SiXu, Yang KaiJing, Zhang Wan, Gao Xue, Zhang ShuYing
Affiliated Zhongshan Hospital of Dalian university, Dalian, 116001, China.
BMC Cardiovasc Disord. 2025 Jun 7;25(1):437. doi: 10.1186/s12872-025-04890-7.
Endothelial cell dysfunction is a fundamental injury of atherosclerosis cardiovascular disease (ASCVD), closely linked to ferroptosis, which is a novel type of cell death induced by iron-dependent lipid peroxidation. Several clinical trials have suggested that empagliflozin, a selective inhibitor of sodium glucose co-transporter 2, reduces the risk of hospitalization for heart failure and cardiovascular death in patients with type 2 diabetes. However, little is known about the mechanism of EMPA in endothelial cell ferroptosis in ASCVD. The aim of the present study was to evaluate the potential mechanism of EMPA against ferroptosis induced by RAS-selective lethal 3 (RSL3) in endothelial cells.EA.hy926 human umbilical vein endothelial cells were cultured in vitro and were divided into four groups: The Control, RSL3, RSL3 + low-concentration EMPA intervention and RSL3 + high-concentration EMPA intervention groups. Iron-dependent lipid peroxidation was assessed by detecting the fluorescence intensity of ferrous ion (Fe), lipid reactive oxygen species (ROS) and content of malondialdehyde (MDA). The expression of ferroptosis-related genes was assessed by RT-qPCR and western blotting. siRNA nuclear factor erythroid 2-related factor 2 (NRF2) was transfected into EA.hy926 cells to measure the expression of target genes.It was demonstrated that the level of MDA, Fe and lipid ROS was higher in the RSL3-treated group compared with the EMPA intervention group, while EMPA markedly mitigated that effect. In addition, EMPA can reverse the RSL3-induced low expression of glutathione peroxidase 4 (GPX4) and high expression of ACSL4 in endothelial cells, as evidenced by the upregulation of nuclear factor transcription factor nuclear factor, erythroid 2 (NFE2)-related factor 2 and heme oxygenase-1 expression, while siNRF2 transfection impaired the antiferroptosis effect of EMPA. The present study indicated that EMPA may inhibit RSL3-induced ferroptosis in endothelial cells by activating the NRF2/heme oxygenase 1 gene pathway.
内皮细胞功能障碍是动脉粥样硬化性心血管疾病（ASCVD）的一种基本损伤，与铁死亡密切相关，铁死亡是一种由铁依赖性脂质过氧化诱导的新型细胞死亡。几项临床试验表明，钠-葡萄糖协同转运蛋白2的选择性抑制剂恩格列净可降低2型糖尿病患者因心力衰竭住院的风险和心血管死亡风险。然而，关于恩格列净在ASCVD内皮细胞铁死亡中的作用机制知之甚少。本研究的目的是评估恩格列净在内皮细胞中对抗RAS选择性致死3（RSL3）诱导的铁死亡的潜在机制。将EA.hy926人脐静脉内皮细胞进行体外培养，并分为四组：对照组、RSL3组、RSL3+低浓度恩格列净干预组和RSL3+高浓度恩格列净干预组。通过检测亚铁离子（Fe）、脂质活性氧（ROS）的荧光强度和丙二醛（MDA）含量来评估铁依赖性脂质过氧化。通过RT-qPCR和蛋白质免疫印迹法评估铁死亡相关基因的表达。将小干扰RNA核因子红细胞2相关因子2（NRF2）转染到EA.hy926细胞中以检测靶基因的表达。结果表明，与恩格列净干预组相比，RSL3处理组的MDA、Fe和脂质ROS水平更高，而恩格列净显著减轻了这种影响。此外，恩格列净可以逆转RSL3诱导的内皮细胞中谷胱甘肽过氧化物酶4（GPX4）低表达和酰基辅酶A合成酶长链家族成员4（ACSL4）高表达，核因子转录因子核因子红细胞2（NFE2）相关因子2和血红素加氧酶-1表达上调证明了这一点，而小干扰RNA NRF2转染削弱了恩格列净的抗铁死亡作用。本研究表明，恩格列净可能通过激活NRF2/血红素加氧酶1基因途径抑制RSL3诱导的内皮细胞铁死亡。
