Ding Hongdou, Xu Xinnan, Zhu Yaoyao, Ling Xinyu, Xu Li
Department of Thoracic Surgery, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, China.
Department of Radiation Oncology, Shanghai Pulmonary Hospital, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, China.
Cell Prolif. 2025 Sep;58(9):e70032. doi: 10.1111/cpr.70032. Epub 2025 Apr 20.
This paper discussed the role of AlkB homologue 5 (Alkbh5) in the progression of lipopolysaccharide (LPS)-induced acute lung injury (ALI). LPS-induced ALI models were established in Alkbh5 knockout (KO) and knock-in (KI) mice. The m6A levels in lung tissues were analysed using m6A dot assays. The lung injury was analysed by determining ALI-related markers and histological staining. Mouse MLE12 cells were exposed to LPS for in vitro experiments, and the influence of Alkbh5 on cell viability, apoptosis and reactive oxygen species (ROS) production was analysed. RNA-seq analysis was performed to analyse gene changes upon Alkbh5 deficiency. Functions of the Alkbh5-C-C motif chemokine ligand 1 (Ccl1) cascade in ALI were further verified using the Alkbh5 antagonist DDO-2728 and a recombinant protein of Ccl1 (mCcl1). Alkbh5 was upregulated in lung tissues following LPS exposure. Alkbh5 knockout in mice mitigated LPS-induced lung injury, as indicated by reduced serum levels of lung injury markers and reduced immune cell infiltration, fibrosis and apoptosis. Conversely, Alkbh5 overexpression in mice resulted in reverse trends. In vitro, Alkbh5 knockdown in MLE12 cells enhanced cell viability while reducing cell apoptosis and ROS production. Mechanistically, Alkbh5 was found to bind to and destabilise Ccl1 mRNA, leading to increased Treg recruitment. Treatment with DDO-2728 or mCcl1 in mice increased Treg infiltration, thus improving lung tissue pathology and reducing lung injury. This study suggests that Alkbh5 is implicated in ALI progression by reducing Ccl1-mediated Treg recruitment, making it a promising target for ALI management.
本文探讨了 AlkB 同源物 5（Alkbh5）在脂多糖（LPS）诱导的急性肺损伤（ALI）进展中的作用。在 Alkbh5 基因敲除（KO）和基因敲入（KI）小鼠中建立了 LPS 诱导的 ALI 模型。使用 m6A 斑点分析法分析肺组织中的 m6A 水平。通过测定 ALI 相关标志物和组织学染色来分析肺损伤。将小鼠 MLE12 细胞暴露于 LPS 进行体外实验，分析 Alkbh5 对细胞活力、凋亡和活性氧（ROS）产生的影响。进行 RNA 测序分析以分析 Alkbh5 缺乏时的基因变化。使用 Alkbh5 拮抗剂 DDO-2728 和 Ccl1 重组蛋白（mCcl1）进一步验证 Alkbh5-C-C 基序趋化因子配体 1（Ccl1）级联在 ALI 中的功能。LPS 暴露后肺组织中 Alkbh5 上调。小鼠中 Alkbh5 基因敲除减轻了 LPS 诱导的肺损伤，表现为肺损伤标志物血清水平降低以及免疫细胞浸润、纤维化和凋亡减少。相反，小鼠中 Alkbh5 过表达导致相反的趋势。在体外，MLE12 细胞中 Alkbh5 敲低增强了细胞活力，同时减少了细胞凋亡和 ROS 产生。机制上，发现 Alkbh5 与 Ccl1 mRNA 结合并使其不稳定，导致调节性 T 细胞（Treg）募集增加。用 DDO-2728 或 mCcl1 处理小鼠增加了 Treg 浸润，从而改善了肺组织病理学并减轻了肺损伤。本研究表明，Alkbh5 通过减少 Ccl1 介导的 Treg 募集参与 ALI 进展，使其成为 ALI 治疗的一个有前景的靶点。
