Antioxidant potential of CORM-A1 and resveratrol during TNF-α/cycloheximide-induced oxidative stress and apoptosis in murine intestinal epithelial MODE-K cells

• Published in: Toxicology and applied pharmacology Vol. 288; no. 2; pp. 161 - 178
• Main Authors: Babu, Dinesh, Leclercq, Georges, Goossens, Vera, Remijsen, Quinten, Vandenabeele, Peter, Motterlini, Roberto, Lefebvre, Romain A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-10-2015
• Subjects: 60 APPLIED LIFE SCIENCES; Animals; Anti-Inflammatory Agents - pharmacology; ANTIOXIDANTS; Antioxidants - pharmacology; APOPTOSIS; Apoptosis - drug effects; Boranes - pharmacology; CARBON MONOXIDE; Carbonates - pharmacology; Cell Line; CORM-A1; CYCLOHEXIMIDE; Cycloheximide - toxicity; Cytoprotection; DEPOLARIZATION; Epithelial Cells - drug effects; Epithelial Cells - immunology; Epithelial Cells - metabolism; Epithelial Cells - pathology; HYDROGEN PEROXIDE; INFLAMMATION; Intestinal epithelial cells; Intestinal Mucosa - drug effects; Intestinal Mucosa - immunology; Intestinal Mucosa - metabolism; Intestinal Mucosa - pathology; Membrane Potential, Mitochondrial - drug effects; MICE; MITOCHONDRIA; Mitochondria - drug effects; Mitochondria - metabolism; NADPH oxidase; NADPH Oxidases - metabolism; OXIDASES; OXIDATION; Oxidative Stress - drug effects; Oxygen Consumption - drug effects; Resveratrol; Stilbenes - pharmacology; Superoxides - metabolism; TNF-α/CHX; Tumor Necrosis Factor-alpha - toxicity; CORM-A1; Mitochondria; TNF-α/CHX; NADPH oxidase; Intestinal epithelial cells; Resveratrol
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2015.07.007

Targeting excessive production of reactive oxygen species (ROS) could be an effective therapeutic strategy to prevent oxidative stress-associated gastrointestinal inflammation. NADPH oxidase (NOX) and mitochondrial complexes (I and II) are the major sources of ROS production contributing to TNF-α/cycloheximide (CHX)-induced apoptosis in the mouse intestinal epithelial cell line, MODE-K. In the current study, the influence of a polyphenolic compound (resveratrol) and a water-soluble carbon monoxide (CO)-releasing molecule (CORM-A1) on the different sources of TNF-α/CHX-induced ROS production in MODE-K cells was assessed. This was compared with H2O2-, rotenone- or antimycin-A-induced ROS-generating systems. Intracellular total ROS, mitochondrial-derived ROS and mitochondrial superoxide anion (O2−) production levels were assessed. Additionally, the influence on TNF-α/CHX-induced changes in mitochondrial membrane potential (Ψm) and mitochondrial function was studied. In basal conditions, CORM-A1 did not affect intracellular total or mitochondrial ROS levels, while resveratrol increased intracellular total ROS but reduced mitochondrial ROS production. TNF-α/CHX- and H2O2-mediated increase in intracellular total ROS production was reduced by both resveratrol and CORM-A1, whereas only resveratrol attenuated the increase in mitochondrial ROS triggered by TNF-α/CHX. CORM-A1 decreased antimycin-A-induced mitochondrial O2− production without any influence on TNF-α/CHX- and rotenone-induced mitochondrial O2− levels, while resveratrol abolished all three effects. Finally, resveratrol greatly reduced and abolished TNF-α/CHX-induced mitochondrial depolarization and mitochondrial dysfunction, while CORM-A1 only mildly affected these parameters. These data indicate that the cytoprotective effect of resveratrol is predominantly due to mitigation of mitochondrial ROS, while CORM-A1 acts solely on NOX-derived ROS to protect MODE-K cells from TNF-α/CHX-induced cell death. This might explain the more pronounced cytoprotective effect of resveratrol.
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•In MODE-K IECs, TNF-α/CHX induces correlating ROS, mitochondrial O2− and cell death.•CORM-A1 does not influence basal intracellular ROS and mitochondrial O2− levels.•Resveratrol increases basal intracellular ROS but decreases mitochondrial O2− levels.•CORM-A1 acts solely on NOX-derived ROS to protect from cell death by TNF-α/CHX.•Cytoprotection by resveratrol is predominantly due to reduction of mitochondrial O2−.