Quantitative Proteomics Reveals Fh15 as an Antagonist of TLR4 Downregulating the Activation of NF-κB, Inducible Nitric Oxide, Phagosome Signaling Pathways, and Oxidative Stress of LPS-Stimulated Macrophages.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:40725160", "kind": "science-record", "revision": 4, "content_hash": "sha256:4ae33cd74b4967a46f74a974dcbc8ad9169a96a8ea96a19821c069b4e61bba39", "created_at": "2026-11-01T20:30:49Z", "updated_at": "2026-11-01T22:56:54Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Quantitative Proteomics Reveals Fh15 as an Antagonist of TLR4 Downregulating the Activation of NF-κB, Inducible Nitric Oxide, Phagosome Signaling Pathways, and Oxidative Stress of LPS-Stimulated Macrophages.", "doi": "10.3390/ijms26146914", "pmid": "40725160", "authors": ["Albersy Armina-Rodriguez", "Bianca N Valdés Fernandez", "Carlimar Ocasio-Malavé", "Yadira M Cantres Rosario", "Kelvin Carrasquillo Carrión", "Loyda M Meléndez", "Abiel Roche Lima", "Eduardo L Tosado Rodriguez", "Ana M Espino"], "journal": "International journal of molecular sciences", "year": 2025, "abstract": "There is a present need to develop alternative biotherapeutic drugs to mitigate the exacerbated inflammatory immune responses characteristic of sepsis. The potent endotoxin lipopolysaccharide (LPS), a major component of Gram-negative bacterial outer membrane, activates the immune system via Toll-like receptor 4 (TLR4), triggering macrophages and a persistent cascade of inflammatory mediators. Our previous studies have demonstrated that Fh15, a recombinant member of the Fasciola hepatica fatty acid binding protein family, can significantly increase the survival rate by suppressing many inflammatory mediators induced by LPS in a septic shock mouse model. Although Fh15 has been proposed as a TLR4 antagonist, the specific mechanisms underlying its immunomodulatory effect remained unclear. In the present study, we employed a quantitative proteomics approach using tandem mass tag (TMT) followed by LC-MS/MS analysis to identify and quantify differentially expressed proteins that participate in signaling pathways downstream TLR4 of macrophages, which can be dysregulated by Fh15. Data are available via ProteomeXchange with identifier PXD065520. Based on significant fold change (FC) cut-off of 1.5 and p-value ≤ 0.05 criteria, we focused our attention to 114 proteins that were upregulated by LPS and downregulated by Fh15. From these proteins, TNFα, IL-1α, Lck, NOS2, SOD2 and CD36 were selected for validation by Western blot on murine bone marrow-derived macrophages due to their relevant roles in the NF-κB, iNOS, oxidative stress, and phagosome signaling pathways, which are closely associated with sepsis pathogenesis. These results suggest that Fh15 exerts a broad spectrum of action by simultaneously targeting multiple downstream pathways activated by TLR4, thereby modulating various aspects of the inflammatory responses during sepsis.", "claims": [{"id": "claim-method", "statement": "Data were analyzed using one-way ANOVA with multiple comparisons and the Tukey test, employing GraphPad Prism software (version 8).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:40725160", "locator": "sections[20] \"4.11. Statistical Analysis\" paragraph 2", "quote": "Data were analyzed using one-way ANOVA with multiple comparisons and the Tukey test, employing GraphPad Prism software (version 8)."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [92] partner repository with the dataset identifier PXD065520 and 10.6019/PXD065520. 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