There is compelling evidence indicating that ethanol and its oxidative metabolite acetaldehyde can disrupt intestinal barrier function. Apart from the tight junctions, mucins secreted by goblet cells provide an effective barrier. Ethanol has been shown to induce goblet cell injury associated with alterations in mucins glycosylation. However, effects of its most injurious metabolite, acetaldehyde remains largely unknown. This study aimed to assess short-term effects of acetaldehyde(0, 25, 50, 75,100 µM) on functional characteristics of intestinal goblet-like cells (LS174T). Oxidative stress, mitochondrial function, ATP and intra-mitochondrial calcium (Ca2+)were assessed by dichlorofluorescein, methyltetrazolium, and bioluminescence, MitoTracker green and rhod-2 double-labeling, respectively. Membrane integrity and apoptosis were evaluated by measuring lactate dehydrogenase (LDH), caspase 3/7 and cleavage of cytokeratin 18 (CK18). Expression of mucin 2 (MUC2) was determined by cell-based ELISA. Acetaldehyde significantly increased ROS generation and decreased mitochondrial function compared to negative controls ( P < 0.05). In addition, acetaldehyde dose-dependently decreased ATP levels and induced intra-mitochondrial Ca2+ accumulation compared to negative controls (P < 0.05). Furthermore, acetaldehyde induced LDH release and increased caspase 3/7 activity and percentage of cells expressing cleaved CK18 and increased MUC2 protein expression compared to negative controls (P < 0.0001). ATP depletion and LDH release could be largely prevented by the antioxidant N-acetylcysteine, suggesting a pivotal role for oxidative stress. Our data demonstrate that acetaldehyde has distinct oxidant-dependent metabolic and cytotoxic effects on LS174T cells that can lead to induction of cellular apoptosis. These effects may contribute to acetaldehyde-induced intestinal barrier dysfunction and subsequently to liver injury.
- goblet cells
- LS174T cells
- Copyright © 2014, American Journal of Physiology- Gastrointestinal and Liver Physiology