Zinc finger transcription factor ZNF384 mitigates LPS-induced ferroptosis and inflammation in lung epithelial cells by activating SESN2-mediated autophagy

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro Pub Date : 2025-05-13 DOI:10.1016/j.tiv.2025.106073

Lu Shi , Hongxue Fu , Yingting Hao , Chang Liu , Fachun Zhou

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Abstract

Background

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.

Methods

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.

Results

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.

Conclusion

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.

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锌指转录因子ZNF384通过激活sesn2介导的自噬，减轻lps诱导的肺上皮细胞铁凋亡和炎症

背景：铁凋亡是一种不同于细胞凋亡的程序性细胞死亡，可能与败血症引发的急性呼吸窘迫综合征（ARDS）有关。ZNF384是一种转录因子，但其在ARDS中的作用尚不清楚。方法采集脓毒症致ARDS患者和健康对照者的血液样本。用脂多糖（LPS）刺激BEAS-2B细胞模拟脓毒症诱导的肺上皮细胞损伤模型。利用RT-qPCR、western blot和免疫荧光染色分别检测基因和蛋白的表达。采用CCK-8和流式细胞术检测细胞活力和死亡情况。采用ELISA法检测炎症因子和氧化应激标志物。DCFH-DA染色测定细胞内ROS。用铁检测试剂盒测定铁浓度。通过ChIP和荧光素酶报告基因检测确认靶关系。结果znf384和SESN2水平在脓毒症诱导的ARDS患者中下调。过表达ZNF384可减轻lps刺激的BEAS-2B细胞的炎症损伤、铁凋亡和自噬增强。从机制上讲，ZNF384作为SESN2的转录激活因子促进自噬激活。救援实验证实，SESN2的缺失显著逆转了ZNF384在lps诱导的炎症、自噬和铁凋亡中的调节功能。结论ZNF384通过激活sesn2介导的自噬，减轻了lps引发的BEAS-2B细胞铁凋亡和炎症，提示ZNF384是治疗ARDS的新靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Toxicology in Vitro 医学-毒理学

CiteScore

6.50

自引率

3.10%

发文量

181

审稿时长

65 days

期刊介绍： Toxicology in Vitro publishes original research papers and reviews on the application and use of in vitro systems for assessing or predicting the toxic effects of chemicals and elucidating their mechanisms of action. These in vitro techniques include utilizing cell or tissue cultures, isolated cells, tissue slices, subcellular fractions, transgenic cell cultures, and cells from transgenic organisms, as well as in silico modelling. The Journal will focus on investigations that involve the development and validation of new in vitro methods, e.g. for prediction of toxic effects based on traditional and in silico modelling; on the use of methods in high-throughput toxicology and pharmacology; elucidation of mechanisms of toxic action; the application of genomics, transcriptomics and proteomics in toxicology, as well as on comparative studies that characterise the relationship between in vitro and in vivo findings. The Journal strongly encourages the submission of manuscripts that focus on the development of in vitro methods, their practical applications and regulatory use (e.g. in the areas of food components cosmetics, pharmaceuticals, pesticides, and industrial chemicals). Toxicology in Vitro discourages papers that record reporting on toxicological effects from materials, such as plant extracts or herbal medicines, that have not been chemically characterized.