Activated clotting factor X mediates mitochondrial alterations and inflammatory responses via protease-activated receptor signaling in alveolar epithelial cells

• Published in: European journal of pharmacology Vol. 869; p. 172875
• Main Authors: Bukowska, Alicja, Schild, Lorenz, Bornfleth, Philipp, Peter, Daniela, Wiese-Rischke, Cornelia, Gardemann, Andreas, Isermann, Berend, Walles, Thorsten, Goette, Andreas
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.02.2020
• Subjects: Activated clotting factor X; Apoptosis; Human lung epithelial cells; Inflammatory signaling; Mitochondrial dysfunction; Protease-activated receptors; “Biased” effects of GB83; Human lung epithelial cells; Mitochondrial dysfunction; “Biased” effects of GB83; Activated clotting factor X; Inflammatory signaling; Protease-activated receptors; Apoptosis
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2019.172875

There is growing evidence for the contribution of the activated coagulation factor X (FXa) in the development of chronic inflammatory lung diseases. Therefore, we aimed to investigate effects of exogenous FXa on mitochondrial and metabolic function as well as the induction of inflammatory molecules in type II alveolar epithelial cells. Effects of FXa on epithelial cells were investigated in A549 cell line. Activation of extracellular signal-regulated kinase (ERK) and induction of inflammatory molecules were examined by immunoblot and gene expression analysis. Mitochondrial function was assessed by the measurement of oxygen consumption during maximal oxidative phosphorylation and quantitative determination of cardiolipin oxidation. Apoptosis was tested using a caspase 3 antibody. Metabolic activity and lactate dehydrogenase assay were applied for the detection of cellular viability. FXa activated ERK1/2 and induced an increase in the expression of pro-inflammatory cytokines, which was prevented by an inhibitor of FXa, edoxaban, or an inhibitor of protease-activated receptor 1, vorapaxar. Exposure to FXa caused mitochondrial alteration with restricted capacity for ATP generation, which was effectively prevented by edoxaban, vorapaxar and GB83 (inhibitor of protease-activated receptor 2). Of note, exposure to FXa did not initiate apoptosis in epithelial cells. FXa-dependent pro-inflammatory state and impairment of mitochondria did not reach the level of significance in lung epithelial cells. However, these effects might limit regenerative potency of lung epithelial cells, particular under clinical circumstances where lung injury causes exposure to clotting factors.