Itaconic acid alleviates macrophage PANoptosis in sepsis-associated acute lung injury via inhibiting ninjurin-1-mediated plasma membrane rupture.
10.11817/j.issn.1672-7347.2025.250283
- Author:
Mengrui CHEN
1
,
2
,
3
;
Xiaohua TAN
4
,
5
;
Wenjing ZHONG
1
;
Hanxi SHA
1
;
Liying LIANG
1
;
Shaokun LIU
6
,
7
Author Information
1. Department of Physiology, Xiangya School of Basic Medicine Science, Central South University, Changsha
2. chenmr01@
3. com.
4. Experimental Center of Medical Morphology, Xiangya School of Basic Medical Science, Central South University, Changsha
5. tanxiaohua@csu.edu.cn.
6. Department of Pulmonary and Critical Care Medicine, Second Xiangya Hospital, Central South University, Changsha
7. shaokunliu228@csu.edu.cn.
- Publication Type:Journal Article
- Keywords:
PANoptosis;
itaconic acid;
macrophages;
ninjurin-1;
plasma membrane rupture;
sepsis-associated acute lung injury
- MeSH:
Animals;
Acute Lung Injury/pathology*;
Succinates/pharmacology*;
Sepsis/complications*;
Mice, Inbred C57BL;
Male;
Mice;
Macrophages/pathology*;
Cell Membrane/metabolism*;
Lipopolysaccharides;
Hydro-Lyases
- From:
Journal of Central South University(Medical Sciences)
2025;50(6):970-985
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES:Sepsis-associated acute lung injury (S-ALI) is one of the major causes of death in intensive care unit (ICU) patients, yet its mechanisms remain incompletely understood and effective therapies are lacking. Lytic cell death of macrophages is a key driver of the inflammatory cascade in S-ALI. PANoptosis, a newly recognized form of lytic cell death characterized by PANoptosome assembly and activation, involves plasma membrane rupture (PMR) mediated by ninjurin-1 (NINJ1), a recently identified pore-forming protein. Itaconic acid is known for its anti-inflammatory effects, but its role in macrophage PANoptosis during S-ALI is unclear. This study aims to investigate the protective effect of itaconic acid on macrophage PANoptosis in S-ALI to provide new therapeutic insights.
METHODS:Male specific-pathogen-free C57BL/6J mice (6-8 weeks, 18-20 g) received intraperitoneal lipopolysaccharide (LPS) to establish a classical S-ALI model. Western blotting was used to assess PANoptosome-related proteins and enzymes involved in the itaconic acid metabolic pathway, while real-time reverse transcription polymerase chain reaction and metabolomics quantified itaconic acid levels. Primary peritoneal macrophages (PMs) were pretreated with the itaconate derivative 4-octyl itaconate (4-OI) and then exposed to tumor necrosis factor alpha (TNF-α) plus interferon gamma (IFN-γ) to induce PANoptosis. Cell viability was evaluated by cell counting kit-8 (CCK-8) assay. Western blotting was employed to quantify enzymes of the itaconate-metabolic pathway in PANoptotic macrophages, to evaluate the impact of 4-OI on PANoptosome-associated proteins, and to determine NINJ1 abundance in lung tissues from S-ALI mice and in PANoptotic macrophages. Fluorescent dye FM4-64 was used to visualize 4-OI-mediated changes in PMR, whereas immunofluorescence staining mapped the effect of 4-OI on both the expression level and membrane localization of NINJ1 in PANoptotic macrophages. The effect of 4-OI on lactate dehydrogenase (LDH) release in culture supernatants and peripheal blood serum was assessed using a LDH assay kit, and non-denataring polyacylamide gel electrophoresis was used to assess the expression of NINJ1 in S-ALI mouse lung tissues and the impact of 4-OI on the expression of PANoptosis-associated NINJ1 multimeric reflected protein in macropahges.
RESULTS:In S-ALI mouse lungs, PANoptosome components [NOD-like receptor thermal protein domain associated protein 3 (NLRP3), Gasdermin D (GSDMD), Caspase-1, Z-DNA binding protein (ZBP1), and Caspase-3] and phosphorylated mixed lineage kinase domain-like protein (MLKL) S345 were significantly upregulated (all P<0.05), while metabolomics showed compensatory increases in itaconic acid and its key enzymes [aconitate decarboxylase 1 (ACOD1)/immunoresponsive gene 1 (IRG1)]. In macrophages, 4-OI obviously suppressed PANoptosome protein expression, reduced LDH release, restored plasma membrane integrity, and inhibited NINJ1 expression and oligomerization at the membrane (P<0.05).
CONCLUSIONS:Itaconic acid may alleviate macrophage PANoptosis in S-ALI by inhibiting NINJ1-mediated plasma membrane rupture. Targeting NINJ1 or enhancing itaconate pathways may offer a novel therapeutic strategy for S-ALI.
