Compound C1 reduced inflammation and activated autophagy in alveolar macrophages in mice

IF 3.2 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular immunology Pub Date : 2025-03-25 DOI:10.1016/j.molimm.2025.03.010

Qi Sheng , Jie Zhao , Shujun Chen , Jingmin Zeng , Shaogui Wang , Jinping Wang

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引用次数: 0

Abstract

The purpose of this study was to investigate the therapeutic effect of Compound C1 (Comp-C1) on lipopolysaccharide (LPS) -mediated sepsis acute lung injury (SALI) in vitro alveolar macrophage model and its regulatory mechanism. In vitro cultured mouse alveolar macrophages (MH-S) were treated with LPS. The expression and localization of transcription factor EB (TFEB) after LPS stimulation were detected. Then the cells were treated with LPS (1 μg/mL) and Comp-C1 (1 μM) for 24 h. RT-qPCR and Western Blot were used to detect the mRNA expression of inflammatory factors. Western blot was used to detect the expression of TFEB, lysosome-associated membrane protein 1 (LAMP1), P62 and microtubule-associated protein 1 light chain 3B (LC3B). TFEB-EGFP-Hela and mCherry-EGFP-LC3-Hela cells were used to detect the changes of TFEB nuclear expression and intracellular autophagic flux after Comp-C1 administration by immunofluorescence. The results showed that the expression of inflammatory factors was the highest after 1 μg / mL LPS stimulation for 24 hours. At the same time, the expression of TFEB gene and protein decreased after LPS stimulation, and the content of TFEB in cytoplasm and nucleus decreased by separating cytoplasmic and nuclear proteins. The content of LAMP1 decreased, and the expression of autophagy-related proteins reflected the inhibition of autophagy. After treatment with Comp-C1, the inflammatory factors were significantly decreased, the expression of TFEB and LAMP1 was significantly increased, and the expression of autophagy genes in the cells was restored. The up-regulation of TFEB nuclear expression after Comp-C1 administration was determined by TFEB-EGFP-Hela cells, and the recovery of autophagy flux and alveolar macrophage function after Comp-C1 administration was determined by mCherry-EGFP-LC3-Hela cells. Therefore, Comp-C1 can alleviate LPS-induced MH-S autophagy dysfunction and reduce inflammatory response by up-regulating TFEB in mouse alveolar macrophages, suggesting that Comp-C1 can be used as a potential drug for the treatment of SALI

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来源期刊

Molecular immunology 医学-免疫学

CiteScore

6.90

自引率

2.80%

发文量

324

审稿时长

50 days

期刊介绍： Molecular Immunology publishes original articles, reviews and commentaries on all areas of immunology, with a particular focus on description of cellular, biochemical or genetic mechanisms underlying immunological phenomena. Studies on all model organisms, from invertebrates to humans, are suitable. Examples include, but are not restricted to: Infection, autoimmunity, transplantation, immunodeficiencies, inflammation and tumor immunology Mechanisms of induction, regulation and termination of innate and adaptive immunity Intercellular communication, cooperation and regulation Intracellular mechanisms of immunity (endocytosis, protein trafficking, pathogen recognition, antigen presentation, etc) Mechanisms of action of the cells and molecules of the immune system Structural analysis Development of the immune system Comparative immunology and evolution of the immune system "Omics" studies and bioinformatics Vaccines, biotechnology and therapeutic manipulation of the immune system (therapeutic antibodies, cytokines, cellular therapies, etc) Technical developments.