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Stimulation of mitochondrial superoxide in lung epithelial cells using a novel stretching device (869.5)
Author(s) -
Rapalo Gabriel,
Roan Esra,
Schwingshakl Andreas,
Eckstein Eugene,
Waters Christopher
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.869.5
Subject(s) - superoxide , microbiology and biotechnology , cytochalasin d , reactive oxygen species , chemistry , mitochondrion , a549 cell , oxidative stress , mitochondrial ros , proinflammatory cytokine , biophysics , cell , biology , cytoskeleton , immunology , inflammation , biochemistry , enzyme
Mechanical ventilation of patients with acute lung injury can cause further damage to the lung epithelium, leading to ventilator‐associated lung injury. We and others have previously demonstrated that cyclic stretch of bronchial and alveolar epithelial cells stimulates increased production of reactive oxygen species (ROS) and other proinflammatory cytokines, but the mechanism is not completely understood. To investigate, we designed a motor‐driven device to apply linear strains up to 20% on cells grown on an elastic substrate while imaging under a confocal microscope. We hypothesized that distention of the lung epithelium triggers an oxidative stress response of mitochondrial superoxide through deformation of the mitochondria and actin filaments. Cultures of an immortalized human airway epithelial cell line (16HBE) and human alveolar type II cell line (A549) were tonically stretched while the production of mitochondrial superoxide was measured using MitoSox. Stretch in 16HBE cells caused a significant increase in production of superoxide, while treatment with Tiron or Cytochalasin D both prevented the stretch‐induced increase in superoxide. A549 cells did not show a significant increase in production of superoxide compared to treatment with the inhibitors. A single tonic deformation of the mitochondria via actin filaments directly stimulated the release of superoxide. Grant Funding Source : Supported by NIH grant HL094366 (CMW)