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Lipopolysaccharide induces apoptotic insults to human alveolar epithelial A549 cells through reactive oxygen species-mediated activation of an intrinsic mitochondrion-dependent pathway

  • Molecular Toxicology
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Abstract

Alveolar type II epithelial cells can regulate immune responses to sepsis-induced acute lung injury. Lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria, can cause septic shock. This study was designed to evaluate the cytotoxic effects of LPS on human alveolar epithelial A549 cells and its possible molecular mechanisms. Exposure of A549 cells to LPS decreased cell viability in concentration- and time-dependent manners. In parallel, LPS concentration- and time-dependently induced apoptosis of A549 cells. Meanwhile, LPS only at a high concentration of 10 μg/ml caused mildly necrotic insults to A549 cells. In terms of the mechanism, exposure of A549 cells to LPS increased the levels of cellular nitric oxide and reactive oxygen species (ROS). Pretreatment with N-acetylcysteine (NAC), an antioxidant, significantly lowered LPS-caused enhancement of intracellular ROS in A549 cells and simultaneously attenuated the apoptotic insults. Sequentially, treatment of A549 cells with LPS caused significant decreases in the mitochondrial membrane potential and biosynthesis of adenosine triphosphate. In succession, LPS triggered the release of cytochrome c from the mitochondria to the cytoplasm. Activities of caspase-9 and caspase-6 were subsequently augmented following LPS administration. Consequently, exposure of A549 cells induced DNA fragmentation in a time-dependent manner. Pretreatment of A549 cells with NAC significantly ameliorated LPS-caused alterations in caspase-9 activation and DNA damage. Therefore, this study shows that LPS specifically induces apoptotic insults to human alveolar epithelial cells through ROS-mediated activation of the intrinsic mitochondrion–cytochrome c-caspase protease mechanism.

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Acknowledgments

This study was supported by the Department of Health, Taipei City Hospital (94002-62-005 and 95001-62-017), Taipei, Taiwan. The authors express their gratitude to Ms. Yi-Ling Lin for technical support and data collection during the experiments.

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Authors and Affiliations

  1. Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan

    Chi-Yuan Chuang & Ta-Liang Chen

  2. Cell Physiology and Molecular Image Research, Wan-Fang Medical Center, Taipei Medical University, Taipei, Taiwan

    Yu-Tyng Tai, Tyng-Guey Chen & Ruei-Ming Chen

  3. Department of Pulmonology, Division of Internal Medicine, Ren-Ai Branch, Taipei City Hospital, Taipei, Taiwan

    Chi-Yuan Chuang

  4. Department of Anesthesiology, Taipei Medical University Hospital, Taipei, Taiwan

    Ta-Liang Chen

  5. Department of Anesthesiology, Shuang Ho Hospital, Taipei, Taiwan

    Yih-Giun Cherng

  6. Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, 250 Wu-Xing St, Taipei, 110, Taiwan

    Ruei-Ming Chen

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  1. Chi-Yuan Chuang
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  2. Ta-Liang Chen
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  3. Yih-Giun Cherng
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  4. Yu-Tyng Tai
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Corresponding author

Correspondence to Ruei-Ming Chen.

Additional information

Chi-Yuan Chuang and Ta-Liang Chen contributed equally to this work.

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Chuang, CY., Chen, TL., Cherng, YG. et al. Lipopolysaccharide induces apoptotic insults to human alveolar epithelial A549 cells through reactive oxygen species-mediated activation of an intrinsic mitochondrion-dependent pathway. Arch Toxicol 85, 209–218 (2011). https://doi.org/10.1007/s00204-010-0585-x

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  • DOI: https://doi.org/10.1007/s00204-010-0585-x

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