Cytotoxicity and metabolic stress induced by acetaldehyde in human intestinal LS174T goblet-like cells

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 mucin glycosylation. However, effects of its most injurious metabolite acetaldehyde remain 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 intramitochondrial calcium (Ca2+) were assessed by dichlorofluorescein, methyltetrazolium, and bioluminescence, MitoTracker green and rhod-2 double-labeling. 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 reactive oxygen species generation and decreased mitochondrial function compared with negative controls ( P < 0.05). In addition, acetaldehyde dose-dependently decreased ATP levels and induced intramitochondrial Ca2+ accumulation compared with negative controls ( P < 0.05). Furthermore, acetaldehyde induced LDH release and increased caspase3/7 activity and percentage of cells expressing cleaved CK18 and increased MUC2 protein expression compared with 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.