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Lipopolysaccharide Activates Toll-Like Receptor 4 and Prevents Cardiac Fibroblast-to-Myofibroblast Differentiation

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Abstract

Bacterial lipopolysaccharide (LPS) is a known ligand of Toll-like receptor 4 (TLR4) which is expressed in cardiac fibroblasts (CF). Differentiation of CF to cardiac myofibroblasts (CMF) is induced by transforming growth factor-β1 (TGF-β1), increasing alpha-smooth muscle actin (α-SMA) expression. In endothelial cells, an antagonist effect between LPS-induced signaling and canonical TGF-β1 signaling was described; however, it has not been studied whether in CF and CMF the expression of α-SMA induced by TGF-β1 is antagonized by LPS and the mechanism involved. In adult rat CF and CMF, α-SMA, ERK1/2, Akt, NF-κβ, Smad3, and Smad7 protein levels were determined by western blot, TGF-β isoforms by ELISA, and α-SMA stress fibers by immunocytochemistry. CF and CMF secrete the three TGF-β isoforms, and the secretion levels of TGF-β2 was affected by LPS treatment. In CF, LPS treatment decreased the protein levels of α-SMA, and this effect was prevented by TAK-242 (TLR4 inhibitor) and LY294002 (Akt inhibitor), but not by BAY 11-7082 (NF-κβ inhibitor) and PD98059 (ERK1/2 inhibitor). TGF-β1 increased α-SMA protein levels in CF, and LPS prevented partially this effect. In addition, in CMF α-SMA protein levels were decreased by LPS treatment, which was abolished by TAK-242. Finally, in CF LPS decreased the p-Smad3 phosphorylation and increased the Smad7 protein levels. LPS treatment prevents the CF-to-CMF differentiation and reverses the CMF phenotype induced by TGF-β1, through decreasing p-Smad3 and increasing Smad7 protein levels.

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Acknowledgements

This work was supported by FONDECYT (Grant 1130300 to G. Díaz-Araya) and CONICYT (Grant 63130233 to S Bolivar).

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Authors and Affiliations

  1. Advanced Center for Chronic Diseases (ACCDiS), Faculty of Chemical and Pharmaceutical Sciences and Faculty of Medicine, University of Chile, 8380492, Santiago, Chile

    Roxana Santana, Rodolfo Landaeta, Pía Boza, Claudio Humeres, Raúl Vivar, Claudia Muñoz, Viviana Pardo, Samuel Fernandez, Renatto Anfossi & Guillermo Diaz-Araya

  2. Department of Pharmacological and Toxicological Chemistry, Faculty of Chemical and Pharmaceutical Sciences, University of Chile, Santiago, Chile

    Samir Bolívar, Roxana Santana, Rodolfo Landaeta, Pía Boza, Claudio Humeres, Raúl Vivar, Claudia Muñoz, Viviana Pardo, Samuel Fernandez, Renatto Anfossi & Guillermo Diaz-Araya

  3. Center of Medical Investigations, Catholic University of Chile, Santiago, Chile

    Pedro Ayala

  4. Faculty of Chemistry and Pharmacy, Atlantic University, Barranquilla, Colombia

    Samir Bolívar

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  1. Samir Bolívar
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Bolívar, S., Santana, R., Ayala, P. et al. Lipopolysaccharide Activates Toll-Like Receptor 4 and Prevents Cardiac Fibroblast-to-Myofibroblast Differentiation. Cardiovasc Toxicol 17, 458–470 (2017). https://doi.org/10.1007/s12012-017-9404-4

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