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 FM_4-64 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.
Animals ; Acute Lung Injury/pathology* ; Succinates/pharmacology* ; Sepsis/complications* ; Mice, Inbred C57BL ; Male ; Mice ; Macrophages/pathology* ; Cell Membrane/metabolism* ; Lipopolysaccharides ; Hydro-Lyases

Objective: To understand the status and related knowledge requirements of health emergency literacy among college students in Shaanxi Province, so as to provide the basis for improving college students health emergency literacy. Methods: A total of 2 723 students from 18 colleges and universities in Shaanxi Province were selected by multi stage random sampling and simple random sampling methods in November 2023, and the survey of health literacy in emergency and knowledge requirements of health emergency literacy was conducted. Statistical analysis was carried out by using χ 2 test, Wilcoxon rank sum test, Kruskal-Wallis H test and Logistic regression analysis. Results: About 28.98% of the surveyed college students had a high level of health emergency literacy, which varied by students whether being only one child, whether having left behind experience, with different personality types, whether being student cadres, and with different frequencies of community or social activities ( χ 2=9.15, 7.90, 32.73, 16.29 , 120.25, P <0.05). The equivalence scores of the four dimensions of health emergency literacy from high to low were poisoning and nuclear and radiation (0.84), medical rescue (0.83), infectious disease (0.82), and basic knowledge and behavior ( 0.77 ). Logistic regression analysis found that college students with left-behind experience were negatively correlated with health emergency literacy and its four dimensions ( OR =0.74, 0.72, 0.80, 0.80, 0.83), while personality type (rational type), community or social activity frequency were positively correlated with the cognitive levels of health emergency literacy and its four dimensions among college students ( OR =1.57, 1.50, 1.33, 1.27, 1.38)( P <0.05). There was a higher level of basic knowledge and behavioral cognition among only child college students ( OR =3.73), and female students had a higher level of health emergency literacy, as well as awareness of infectious disease outbreaks and medical rescue ( OR =1.21, 1.28, 1.21)( P <0.05). The radar map showed that the level of health emergency literacy was positive development radar map. About 67.68 % of the students had a high willingness to acquire health emergency literacy knowledge, and the demand for basic health emergency knowledge and behavioral knowledge was the highest (52.37%). Conclusions College students have insufficient health emergency literacy, but they have the highest demand for health emergency. Publicity and education should be strengthened for students with left behind experience, irrational type, and low frequency of community or social activities.