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LCN2基因敲除可能通过抑制TLR4/NF-κB信号通路改善脓毒症小鼠肺损伤和铁死亡

王奕人^1,2, 于梦洁^1,2, 杜若丽^1,2, 贺玉莹^2,3, 张贻欣⁴, 王佳慧², 叶红伟^1,2, 高琴^1,2,*

¹蚌埠医科大学生理学教研室，蚌埠 233030；²蚌埠医科大学心脑血管疾病基础与临床重点实验室，蚌埠 233030；³蚌埠医科大学生命科学学院，蚌埠 233030；⁴蚌埠医科大学临床医学院，蚌埠 233030

摘要

本研究旨在探讨脂质运载蛋白 2 (lipocalin-2, LCN2)基因敲除(LCN2KO)对盲肠结扎与穿孔(cecal ligation andperforation, CLP)诱导的脓毒症小鼠肺损伤和铁死亡的作用，进一步明确Toll 样受体4 (Toll-like receptor 4, TLR4)/核因子-κB(nuclear factor κB, NF-κB)通路在其中的可能作用。用LCN2KO 小鼠建立CLP 模型，用ELISA 检测小鼠血清中白介素6(interleukin-6, IL-6)和IL-1β 含量，用试剂盒检测肺组织丙二醛(malondialdehyde, MDA)和谷胱甘肽(glutathione, GSH)含量，用DHE荧光探针检测肺组织中活性氧(reactive oxygen species, ROS)水平，用Western blot 检测肺组织中铁死亡相关蛋白的表达水平。结果显示，相对野生型(wild type, WT)-Sham 组，WT-CLP组小鼠肺组织明显受损，血清IL-6 和IL-1β 含量升高，肺组织MDA含量增加，GSH含量降低，ROS水平显著升高，LCN2 和TLR4 蛋白表达水平显著上调，p-P65/P65 比值显著升高，铁死亡抑制蛋白1 (ferroptosis suppressor protein 1, FSP1)和谷胱甘肽过氧化物酶4 (glutathione peroxidase 4, GPX4)蛋白表达水平显著下调。LCN2KO可以逆转脓毒症小鼠的上述变化。TLR4 抑制剂TAK-242 和NF-κB抑制剂PDTC均可下调脓毒症小鼠肺组织中LCN2 蛋白表达水平；与LCN2KO的作用相似，二者均可逆转脓毒症诱导的铁死亡和肺损伤。以上结果提示，LCN2KO可能通过调控TLR4/NF-κB信号通路减轻铁死亡，从而改善脓毒症小鼠肺损伤。

关键词： 脂质运载蛋白2; 脓毒症肺损伤; 铁死亡; TLR4/NF-κB信号通路; 炎症

Knockout of LCN2 attenuates lung injury and ferroptosis in septic mice likely through inhibiting the TLR4/NF-κB signaling pathway

WANG Yi-Ren^1,2, YU Meng-Jie^1,2, DU Ruo-Li^1,2, HE Yu-Ying^2,3, ZHANG Yi-Xin⁴, WANG Jia-Hui², YE Hong- Wei^1,2, GAO Qin^1,2,*

¹Department of Physiology, Bengbu Medical University, Bengbu 233030, China；²Key Laboratory of Basic and Clinical Cardiovascular Diseases, Bengbu Medical University, Bengbu 233030, China；³School of Life Sciences, Bengbu Medical University, Bengbu 233030, China；⁴School of Clinical Medicine, Bengbu Medical University, Bengbu 233030, China

Abstract

This study aimed to investigate the effects of lipocalin-2 (LCN2) gene knockout (LCN2KO) on cecal ligation and perforation (CLP) -induced lung injury and ferroptosis in septic mice, and further to clarify the potential role of the Toll-like receptor 4 (TLR4)/nuclear factor κB (NF-κB) pathway. A CLP model was established using LCN2KO mice. The levels of serum interleukin-6 (IL- 6) and IL-1β were measured by ELISA. The contents of malondialdehyde (MDA) and glutathione (GSH) in lung tissue were determined using the corresponding assay kits. Reactive oxygen species (ROS) levels in lung tissue were detected using dihydroethidium (DHE) fluorescent probe. The expression levels of ferroptosis-related proteins in lung tissue were assessed by Western blot. The results showed that, compared to wild-type (WT)-Sham group, WT-CLP group exhibited significant lung tissue damage, elevated serum IL-6 and IL-1β levels, increased MDA content and ROS level, decreased GSH content, up-regulated protein expression levels of LCN2 and TLR4, increased p-P65/P65 ratio, and significantly down-regulated protein expression levels of ferroptosis suppressor protein 1 (FSP1) and glutathione peroxidase 4 (GPX4). LCN2KO reversed these changes induced by CLP. Both TLR4 inhibitor TAK- 242 and NF-κB inhibitor PDTC down-regulated lung LCN2 protein expression, reversed sepsis-induced ferroptosis and lung injury, which is similar to the role of LCN2KO. These results suggest that LCN2KO may ameliorate lung injury in sepsis-induced mouse model by attenuating ferroptosis through modulating the TLR4/NF-κB signaling pathway.

Key words: lipocalin-2; sepsis-induced lung injury; ferroptosis; TLR4/NF-κB signaling pathway; inflammation

收稿日期：　　录用日期：

通讯作者：高琴　　E-mail:

DOI: 10.13294/j.aps.2025.0070

引用本文：

王奕人, 于梦洁, 杜若丽, 贺玉莹, 张贻欣, 王佳慧, 叶红伟, 高琴. LCN2基因敲除可能通过抑制TLR4/NF-κB信号通路改善脓毒症小鼠肺损伤和铁死亡[J]. 生理学报 2026; 78 (2): 452-462.

WANG Yi-Ren, YU Meng-Jie, DU Ruo-Li, HE Yu-Ying, ZHANG Yi-Xin, WANG Jia-Hui, YE Hong- Wei, GAO Qin. Knockout of LCN2 attenuates lung injury and ferroptosis in septic mice likely through inhibiting the TLR4/NF-κB signaling pathway. Acta Physiol Sin 2026; 78 (2): 452-462 (in Chinese with English abstract).