Zhang Wenlong, Wang Xinhua, Ma Chenhui, Liang Bao, Ma Lihong, Wang Yan, Lin Yuanjie, Han Shuguang
Department of Respiratory and Critical Care Medicine, Wuxi No. 2 People's Hospital, Jiangnan University Medical Center, No. 68 Zhongshan Road, Wuxi City, Jiangsu Province 214000, China.
Department of Respiratory and Critical Care Medicine, Wuxi No. 2 People's Hospital, Jiangnan University Medical Center, No. 68 Zhongshan Road, Wuxi City, Jiangsu Province 214000, China.
Respir Physiol Neurobiol. 2025 Jan;331:104346. doi: 10.1016/j.resp.2024.104346. Epub 2024 Sep 11.
Acute lung injury (ALI) is a life-threatening condition characterized by excessive pulmonary inflammation, yet its precise pathophysiology remains elusive. Pyroptosis, a programmed cell death mechanism controlled by gasdermin D (GSDMD), has been linked to the etiology of ALI. This study investigated the regulatory functions of the transcription factor E-twenty-six variant gene 5 (ETV5) and GSDMD in ALI.
Lipopolysaccharide (LPS) was used to treat BEAS-2B cells (50 mmol/mL) and establish an LPS-induced mouse model of ALI (by intratracheal administration, 3 mg/kg). Protein-protein docking, immunofluorescence analysis, western blotting, real-time quantitative polymerase chain reaction, and dual-luciferase reporter gene assay were used to examine ETV5-mediated negative feedback regulation of GSDMD and its effects on pyroptosis and ALI.
Our results showed that the physiological function of ETV5 was reduced by its downregulated expression, which impeded its nuclear translocation in ALI mice. Increased pyroptosis and enhanced production of inflammatory cytokines were associated with LPS-induced ALI. ETV5 overexpression in LPS-treated BEAS-2B cells decreased the expression of total and membrane-bound GSDMD, negatively regulated GSDMD, and prevented pyroptosis. The expression of inflammatory cytokines was subsequently reduced due to this inhibition, which, in turn, reduced ALI. Molecular docking analysis and dual-luciferase reporter gene assay results indicated a direct interaction between ETV5 and GSDMD, which inhibited GSDMD production.
Our results indicate that ETV5 inhibits pyroptosis, decreases the expression of inflammatory cytokines, and negatively regulates GSDMD expression to ameliorate ALI symptoms.
急性肺损伤（ALI）是一种危及生命的疾病，其特征为肺部过度炎症，但确切的病理生理学机制仍不清楚。焦亡是一种由gasdermin D（GSDMD）控制的程序性细胞死亡机制，与ALI的病因有关。本研究调查了转录因子E-26变异基因5（ETV5）和GSDMD在ALI中的调节功能。
使用脂多糖（LPS）处理BEAS-2B细胞（50 mmol/mL），并建立LPS诱导的ALI小鼠模型（通过气管内给药，3 mg/kg）。采用蛋白质-蛋白质对接、免疫荧光分析、蛋白质印迹、实时定量聚合酶链反应和双荧光素酶报告基因检测，以研究ETV5介导的对GSDMD的负反馈调节及其对焦亡和ALI的影响。
我们的结果表明，ETV5的表达下调导致其生理功能降低，这阻碍了其在ALI小鼠中的核转位。焦亡增加和炎性细胞因子产生增强与LPS诱导的ALI相关。在LPS处理的BEAS-2B细胞中过表达ETV5可降低总GSDMD和膜结合GSDMD的表达，对GSDMD进行负调节，并防止焦亡。由于这种抑制作用，炎性细胞因子的表达随后降低，进而减轻了ALI。分子对接分析和双荧光素酶报告基因检测结果表明ETV5与GSDMD之间存在直接相互作用，从而抑制了GSDMD的产生。
我们的结果表明，ETV5可抑制焦亡，降低炎性细胞因子的表达，并对GSDMD表达进行负调节，从而改善ALI症状。
