Proteomic analysis and identification reveal the anti-inflammatory mechanism of clofazimine on lipopolysaccharide-induced acute lung injury in mice

Abstract

Background and objective Acute lung injury (ALI)/ acute respiratory distress syndrome (ARDS) was increasingly recognized as one of the most severe acute hyperimmune response of coronavirus disease 2019 (COVID-19). Clofazimine (CFZ) has attracted attention due to its anti-inflammatory property in immune diseases as well as infectious diseases. However, the role and potential molecular mechanism of CFZ in anti-inflammatory responses remain unclear. Methods We analyze the protein expression profiles of CFZ and LPS from Raw264.7 macrophages using quantitative proteomics. Next, the protective effect of CFZ on LPS-induced inflammatory model is assessed, and its underlying mechanism is validated by molecular biology analysis. Results LC–MS/MS-based shotgun proteomics analysis identified 4746 (LPS) and 4766 (CFZ) proteins with quantitative information. The key proteins and their critical signal transduction pathways including TLR4/NF-κB/HIF-1α signaling was highlighted, which was involved in multiple inflammatory processes. A further analysis of molecular biology revealed that CFZ could significantly inhibit the proliferation of Raw264.7 macrophages, decrease the levels of TNF-α and IL-1β, alleviate lung histological changes and pulmonary edema, improve the survival rate, and down-regulate TLR4/NF-κB/HIF-1α signaling in LPS model. Conclusion This study can provide significant insight into the proteomics-guided pharmacological mechanism study of CFZ and suggest potential therapeutic strategies for infectious disease. Supplementary Information The online version contains supplementary material available at 10.1007/s00011-022-01623-w.

pepDESC: A Method for the Detection of Differentially Expressed Proteins for Mass Spectrometry-Based Single-Cell Proteomics Using Peptide-level Information

“…The spike-in datasets D2 and D3 contain seven and four technical replicates in each sample group, respectively. The design of group size considered the current sample size of label-free quantitative MS-based proteomics experiment ( 3 , 16 , 17 ). The sample preparation of dataset D2, the database searching of the three datasets, and the design of the dataset D1 are explicated herein with detailed information presented below.…”

Molecular mechanisms of resistance and treatment efficacy of clofazimine and bedaquiline against Mycobacterium tuberculosis

“…CFZ, a riminophenazine antibiotic, was initially discovered in Dublin in the 1950s’ and used in the treatment of leprosy caused by M. leprae ( 24 , 25 ). CFZ possesses both anti-inflammatory, pro-oxidative, and anti-mycobacterial properties ( 26 , 27 ). Recently, this drug has gained attention once more as a substitute for treating Mtb infections ( 24 , 28 ).…”

“…Recently, CFZ has garnered attention in order to its ability anti-inflammatory properties in both infectious diseases and immune diseases including Crohn disease, discoid lupus erythematosus, ulcerative colitis, chronic lymphocytic leukemia, etc. ( 26 ). Notably, the pandemic status of the coronavirus disease of 2019 (COVID-19) was announced in March 2020.…”

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