南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (8): 1779-1789.doi: 10.12122/j.issn.1673-4254.2026.08.06

• • 上一篇    

敲除脂质运载蛋白2通过抑制NF-κB通路改善脓毒症急性肺损伤小鼠血管内皮屏障功能

于梦洁1,2(), 赵安博1,2, 郝艺璇1,2, 王奕人1,2, 刘月娴3, 叶红伟1,2, 高琴1,2()   

  1. 1.蚌埠医科大学，生理学教研室，安徽 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，安徽 蚌埠 233030
    3.蚌埠医科大学，临床医学院，安徽 蚌埠 233030
  • 收稿日期:2025-12-01 出版日期:2026-08-20 发布日期:2026-08-01
  • 通讯作者: 高琴 E-mail:ymj11711@163.com;bbmcgq@126.com
  • 作者简介:于梦洁，在读硕士研究生，E-mail: ymj11711@163.com
  • 基金资助:
    安徽省高等学校科学研究项目重大项目(2022AH040215);安徽省高校优秀科研创新团队资助项目(2022AH010083);蚌埠医科大学“512”人才项目(by51201102)

LCN2 knockout protects endothelial barrier function in mice with sepsis-induced acute lung injury by regulating the NF-κB signaling pathway

Mengjie YU1,2(), Anbo ZHAO1,2, Yixuan HAO1,2, Yiren WANG1,2, Yuexian LIU3, Hongwei YE1,2, Qin GAO1,2()   

  1. 1.Department of Physiology, Bengbu Medical University, Bengbu 233030, China
    2.Key Laboratory of Basic and Clinical Research on Cardiovascular and Cerebrovascular Diseases, Bengbu Medical University, Bengbu 233030, China
    3.Clinical Medical College, Bengbu Medical University, Bengbu 233030, China
  • Received:2025-12-01 Online:2026-08-20 Published:2026-08-01
  • Contact: Qin GAO E-mail:ymj11711@163.com;bbmcgq@126.com

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摘要：

目的 探讨敲除脂质运载蛋白2（LCN2）基因对脓毒症小鼠急性肺损伤及血管内皮功能障碍的影响，明确核因子-κB（NF-κB）通路在其中的可能作用。 方法 选用20只LCN2基因敲除（LCN2KO）小鼠和60只C57BL/6小鼠，通过盲肠结扎与穿孔（CLP）术构建脓毒症模型，分为WT-Sham组、Sham+PDTC组、Sham+Bay 11-7082组、WT-CLP组、LCN2KO-Sham组，LCN2KO-CLP组、CLP+NF-κB抑制剂PDTC组、CLP+NF-κB抑制剂Bay 11-7082组，10只/组。HE染色观察肺组织病理变化，检测肺湿干比（W/D）、肺指数及伊文思蓝外渗评价肺水肿与血管通透性。ELISA测定血清肿瘤坏死因子-α（TNF-α）及白细胞介素-6（IL-6）水平。免疫荧光检测NF-κB、细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）与CD31共定位情况，TUNEL/CD31双染评估内皮细胞凋亡变化。Western blotting检测LCN2、p-P65/P65、ICAM-1、VCAM-1及血管内皮钙黏附蛋白（VE-cadherin）蛋白表达。免疫共沉淀分析LCN2与P65的潜在相互作用。 结果 WT-CLP组小鼠肺组织结构明显受损，伴随水肿及炎症反应。与WT-CLP组相比，LCN2KO-CLP组及CLP+PDTC组小鼠肺组织损伤减轻，W/D值、肺指数及伊文思蓝外渗量下降，血清TNF-α和IL-6水平降低，肺血管内皮细胞中NF-κB、ICAM-1、VCAM-1与CD31的共定位信号减弱。CLP术后LCN2、p-P65/P65、ICAM-1及VCAM-1表达升高，VE-cadherin表达降低，内皮细胞凋亡增加，LCN2KO及抑制剂干预后相关蛋白表达得到改善，凋亡水平下降。免疫共沉淀结果显示LCN2与P65在肺组织中可能存在相互作用。Bay 11-7082干预后，降低了炎症因子，下调了p-P65及其下游黏附分子表达，恢复VE-cadherin水平。 结论 敲除LCN2可通过抑制NF-κB信号通路减轻脓毒症诱导的急性肺损伤及血管内皮功能障碍。

关键词: 脂质运载蛋白2, 脓毒症肺损伤, 内皮功能障碍, NF-κB信号通路

Abstract:

Objective To investigate the protective effect of lipocalin-2 (LCN2) gene knockout against sepsis-induced acute lung injury (ALI) and vascular endothelial dysfunction in mice and the mediating role of the nuclear factor-κB (NF-κB) signaling pathway. Methods Wild-type (WT) C57BL/6 mice receiving sham operation or cecal ligation and puncture (CLP) to induce sepsis were randomized into 3 subgroups for intraperitoneal injections of saline, PDTC, or Bay 11-7082 (n=10). Twenty LCN2 knockout (LCN2KO) mice were randomized for sham operation or CLP modeling. Pulmonary histopathological changes, lung wet-to-dry (W/D) ratio, lung index, Evans blue extravasation, and serum TNF-α and IL-6 levels in the mice were assessed. The colocalization of NF-κB, ICAM-1, and VCAM-1 with CD31 were analyzed using immunofluorescence staining, and endothelial cell apoptosis was examined with TUNEL/CD31 double staining. Western blotting and co-immunoprecipitation assay were performed to analyze the protein expressions of LCN2, p-P65/P65, ICAM-1, VCAM-1 and VE-cadherin and the interaction between LCN2 and P65. Results The WT mice receiving CLP showed severe lung structural damage with obvious edema and inflammation. In contrast, both the LCN2KO mouse and PDTC-treated WT mouse models of CLP showed milder lung injury with lower W/D ratio, lung index, Evans blue leakage, serum TNF‑α and IL-6 levels and markedly reduced colocalization signals of NF-κB, ICAM-1, and VCAM-1 with CD31. CLP resulted in significantly increased expression levels of LCN2, p-P65/P65, ICAM-1, and VCAM-1, lowered VE-cadherin levels, and enhanced endothelial cell apoptosis, and all these changes were significantly ameliorated in LCN2KO mice and WT mice with NF‑κB inhibitor treatment. Co-immunoprecipitation results suggested the interaction between lung LCN2 and P65. Bay 11-7082 significantly reduced inflammatory cytokine production, downregulated p-P65 expression and its downstream adhesion molecules, and restored VE-cadherin expression. Conclusion LCN2 knockout alleviates sepsis-induced ALI and vascular endothelial dysfunction in mice by inhibiting NF-κB signaling.

Key words: lipocalin-2, sepsis-induced acute lung injury, endothelial dysfunction, nuclear factor-κB signaling pathway