Shi Lu, Fu Hongxue, Hao Yingting, Liu Chang, Zhou Fachun
Department of Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China; Department of Critical Care Medicine, Chongqing Shapingba Traditional Chinese Medicine Hospital, Chongqing 400030, PR China.
Department of Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.
Toxicol In Vitro. 2025 Aug;107:106073. doi: 10.1016/j.tiv.2025.106073. Epub 2025 May 13.
Ferroptosis, a type of programmed cell death distinct from apoptosis, is potentially associated with sepsis-triggered acute respiratory distress syndrome (ARDS). ZNF384 is a transcription factor, but its role in ARDS remain unclear.
Blood samples were collected from sepsis-induced ARDS patients and healthy controls. BEAS-2B cells were stimulated with lipopolysaccharide (LPS) to mimic sepsis-induced damaged lung epithelial cell model. Gene and protein expression were elevated utilizing RT-qPCR, western blot, and immunofluorescent staining, respectively. Cell viability and death were evaluated by CCK-8 and flow cytometry. Inflammatory cytokines and oxidative stress markers were measured using ELISA. Intracellular ROS was determined using DCFH-DA staining. Iron concentration was measured using an iron detection kit. Target relationships were confirmed through ChIP and luciferase reporter assays.
ZNF384 and SESN2 levels were downregulated in sepsis-induced ARDS patients. Overexpression of ZNF384 reduced inflammatory injury, ferroptosis and enhanced autophagy in LPS-stimulated BEAS-2B cells. Mechanistically, ZNF384 served as transcriptional activator of SESN2 to boost autophagy activation. Rescue experiments validated that depletion of SESN2 strikingly reversed the regulatory function of ZNF384 in LPS-induced inflammation, autophagy and ferroptosis.
ZNF384 alleviated LPS-triggered ferroptosis and inflammation in BEAS-2B cells via activating SESN2-mediated autophagy, indicating ZNF384 was a novel target for ARDS treatment.
铁死亡是一种不同于凋亡的程序性细胞死亡，可能与脓毒症引发的急性呼吸窘迫综合征（ARDS）相关。锌指蛋白384（ZNF384）是一种转录因子，但其在ARDS中的作用尚不清楚。
收集脓毒症诱导的ARDS患者和健康对照者的血液样本。用脂多糖（LPS）刺激BEAS-2B细胞以模拟脓毒症诱导的肺上皮细胞损伤模型。分别采用逆转录定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法和免疫荧光染色法检测基因和蛋白表达。通过细胞计数试剂盒-8（CCK-8）法和流式细胞术评估细胞活力和死亡情况。采用酶联免疫吸附测定（ELISA）法检测炎性细胞因子和氧化应激标志物。用2′,7′-二氯二氢荧光素二乙酸酯（DCFH-DA）染色法测定细胞内活性氧（ROS）。使用铁检测试剂盒测量铁浓度。通过染色质免疫沉淀（ChIP）和荧光素酶报告基因检测法确认靶标关系。
脓毒症诱导的ARDS患者中ZNF384和硒代半胱氨酸裂解酶2（SESN2）水平下调。ZNF384过表达可减轻LPS刺激的BEAS-2B细胞的炎性损伤、铁死亡并增强自噬。机制上，ZNF384作为SESN2的转录激活因子促进自噬激活。挽救实验证实，敲低SESN2可显著逆转ZNF384对LPS诱导的炎症、自噬和铁死亡的调节作用。
ZNF384通过激活SESN2介导的自噬减轻LPS诱导的BEAS-2B细胞铁死亡和炎症，表明ZNF384是ARDS治疗中的一个新靶点。
