Zhang Jiayi, He Shuang, Wu Ke, Wei Yifan, Li Chunyu, Zhu Qian, Lian Siyu, Wang Li, Wan Fang, Peng Fuxiang, Zhang Juan, Chen Zongyu, Zhang Xianming
Department of Respiratory and Critical Care Medicine, Affiliated Hospital of Guizhou Medical University, Guiyang, 550004, China.
School of Clinical Medicine, Guizhou Medical University, Guiyang, 550004, China.
Sci Rep. 2025 Jul 1;15(1):21199. doi: 10.1038/s41598-025-04730-w.
Ventilator induced lung injury (VILI) is caused by improper use of mechanical ventilation, and its pathogenesis remains unclear. The aim of this study was to establish animal and cell models of VILI, and to explore the mechanism of miR-125b-5p in alleviating VILI by inhibiting ferroptosis through targeted regulation of Keap1/Nrf2/GPX4 axis. Firstly, ferrostain-1(Fer-1), a ferroptosis inhibitor, was used to confirm that ferroptosis was involved in the progression of VILI. Secondly, overexpression and knockdown of miR-125b-5p were performed to validate its function; Further, mechanistically, miR-125b-5p targets negatively regulated Keap1 to activate Nrf2 and then increased the expression of GPX4, thereby inhibiting the occurrence of ferroptosis. Finally, the rescue experiment shows, overexpression of Keap1 and use of the GPX4 inhibitor RSL3 reversed the miR-125b-5p effect, respectively. Through real-time quantitative polymerase chain reaction (qRT-PCR), western blotting (WB), immunofluorescence (IF), hematoxylin and eosin (H&E), and iron death related factor detection, it was confirmed that, overexpression of miR-125b-5p upregulates ferroptosis inhibitory protein and downregulates ferroptosis promoting protein, leading to alleviation of lung injury. However, overexpression of Keap1 and RSL3 reverses the effect of miR-125b-5p, respectively. Therefore, miR-125b-5p can inhibit ferroptosis and alleviate lung injury in VILI rats by targeting the Keap1/Nrf2/GPX4 axis, miR-125b-5p may be a potential intervention target for VILI.
呼吸机诱导的肺损伤（VILI）是由机械通气使用不当引起的，其发病机制尚不清楚。本研究旨在建立VILI的动物和细胞模型，并通过靶向调控Keap1/Nrf2/GPX4轴抑制铁死亡来探讨miR-125b-5p减轻VILI的机制。首先，使用铁死亡抑制剂铁抑素-1（Fer-1）来证实铁死亡参与了VILI的进展。其次，进行miR-125b-5p的过表达和敲低以验证其功能；此外，从机制上讲，miR-125b-5p靶向负调控Keap1以激活Nrf2，进而增加GPX4的表达，从而抑制铁死亡的发生。最后，挽救实验表明，Keap1的过表达和GPX4抑制剂RSL3的使用分别逆转了miR-125b-5p的作用。通过实时定量聚合酶链反应（qRT-PCR）、蛋白质免疫印迹（WB）、免疫荧光（IF）、苏木精-伊红（H&E）染色以及铁死亡相关因子检测，证实miR-125b-5p的过表达上调了铁死亡抑制蛋白，下调了铁死亡促进蛋白，从而减轻了肺损伤。然而，Keap1的过表达和RSL3分别逆转了miR-125b-5p的作用。因此，miR-125b-5p可通过靶向Keap1/Nrf2/GPX4轴抑制铁死亡并减轻VILI大鼠的肺损伤，miR-125b-5p可能是VILI的一个潜在干预靶点。
