Lactic Acid Is Elevated in Idiopathic Pulmonary Fibrosis and Induces Myofibroblast Differentiation via pH-Dependent Activation of Transforming Growth Factor-β.
ABSTRACT Rationale: Idiopathic pulmonary fibrosis (IPF) is a complex disease for which the pathogenesis is poorly understood. In this study, we identified lactic acid as a metabolite that is elevated in the lung tissue of patients with IPF. Objectives: This study examines the effect of lactic acid on myofibroblast differentiation and pulmonary fibrosis. Methods: We used metabolomic analysis to examine cellular metabolism in lung tissue from patients with IPF and determined the effects of lactic acid and lactate dehydrogenase-5 (LDH5) overexpression on myofibroblast differentiation and transforming growth factor (TGF)-β activation in vitro. Measurements and Main Results: Lactic acid concentrations from healthy and IPF lung tissue were determined by nuclear magnetic resonance spectroscopy; α-smooth muscle actin, calponin, and LDH5 expression were assessed by Western blot of cell culture lysates. Lactic acid and LDH5 were significantly elevated in IPF lung tissue compared with controls. Physiologic concentrations of lactic acid induced myofibroblast differentiation via activation of TGF-β. TGF-β induced expression of LDH5 via hypoxia-inducible factor 1α (HIF1α). Importantly, overexpression of both HIF1α and LDH5 in human lung fibroblasts induced myofibroblast differentiation and synergized with low-dose TGF-β to induce differentiation. Furthermore, inhibition of both HIF1α and LDH5 inhibited TGF-β-induced myofibroblast differentiation. Conclusions: We have identified the metabolite lactic acid as an important mediator of myofibroblast differentiation via a pH-dependent activation of TGF-β. We propose that the metabolic milieu of the lung, and potentially other tissues, is an important driving force behind myofibroblast differentiation and potentially the initiation and progression of fibrotic disorders.
- American Journal of Respiratory and Critical Care Medicine 10/2012; 186(8):701-3. DOI:10.1164/rccm.201208-1491ED · 11.99 Impact Factor
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ABSTRACT: Lung inflammation can result from exposure to multiple types of inflammatory stimuli. Fibroblasts, key structural cells in the lung which are integral to inflammation and wound healing, produce inflammatory mediators following exposure to stimuli such as IL-1β. We and others have shown that the NF-κB member RelB has anti-inflammatory properties in mice. Little is known, however, about the anti-inflammatory role of RelB in human cells and how it functions. MicroRNAs (miRNAs), a novel class of small, non-coding RNAs, can mediate inflammatory signaling pathways including NF-κB through regulation of target gene expression. Our goal was to analyze the anti-inflammatory properties of RelB in human lung fibroblasts. We hypothesized that RelB regulates inflammatory mediator production in lung fibroblasts in part through a mechanism involving miRNAs. To accomplish this, we transfected human lung fibroblasts with a plasmid encoding RelB and siRNA targeting RelB mRNA to overexpress and downregulate RelB, respectively. Interleukin-1β (IL-1β), a powerful proinflammatory stimulus, was used to induce NF-κB-driven inflammatory responses. RelB overexpression reduced IL-1β-induced cyclooxygenase-2 (Cox-2), prostaglandin E2 (PGE2) and cytokine production, and RelB downregulation increased Cox-2 expression and PGE2 production. Further, RelB overexpression increased IL-1β-induced expression of miR-146a, an NF-κB-dependent miRNA with anti-inflammatory properties, while RelB downregulation reduced miR-146a. MiR-146a overexpression ablated the effects of RelB downregulation on IL-1β-induced Cox-2, PGE2 and IL-6 production, suggesting that RelB mediates IL-1β-induced inflammatory mediator production in lung fibroblasts through miR-146a. RelB and miR-146a may therefore be new therapeutic targets in the treatment of lung inflammation caused by various agents and conditions.AJP Lung Cellular and Molecular Physiology 04/2013; DOI:10.1152/ajplung.00352.2012 · 4.04 Impact Factor
- American Journal of Respiratory and Critical Care Medicine 05/2013; 187(9):920-5. DOI:10.1164/rccm.201302-0331UP · 11.99 Impact Factor