In vitro impact of amino acid-derived bacterial metabolites on colonocyte mitochondrial activity, oxidative stress response and DNA integrity

Lucie Armand (1) , Mireille Andriamihaja (1) , Stacy Gellenoncourt (1) , Vincent Bitane (1) , Annaïg Lan (1) , Francois Blachier (1)

1 PNCA - Physiologie de la Nutrition et du Comportement Alimentaire

AbstractBackground4-hydroxyphenylacetic acid (HO-PAA) is produced by intestinal microbiota from L-tyrosine. High concentrations in human fecal water have been associated with cytotoxicity, urging us to test HO-PAA's effects on human colonocytes. We compared these effects with those of phenylacetic acid (PAA), phenol and acetaldehyde, also issued from amino acids fermentation.MethodsHT-29 Glc−/+ human colonocytes were exposed for 24 h to metabolites at concentrations between 350 and 1000 μM for HO-PAA and PAA, 250-1500 μM for phenol and 25-500 μM for acetaldehyde. We evaluated metabolites'cytotoxicity with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide and DNA quantification assays, reactive oxygen species (ROS) production with H2DCF-DA, and DNA damage with the comet assay. We measured cell oxygen consumption and mitochondrial complexes activity by polarography.ResultsAlthough HO-PAA displayed no cytotoxic effect on colonocytes, it decreased mitochondrial complex I activity and oxygen consumption. This was paralleled by an increase in ROS production and DNA alteration. Cells pretreatment with N-acetylcysteine, a ROS scavenger, decreased genotoxic effects of HO-PAA, indicating implication of oxidative stress in HO-PAA's genotoxicity. PAA and phenol did not reproduce these effects, but were cytotoxic towards colonocytes. Last, acetaldehyde displayed no effect in terms of cytotoxicity and mitochondrial metabolic activity, but increased DNA damage.ConclusionsSeveral bacterial metabolites produced from amino acids displayed deleterious effects on human colonocytes, in terms of genotoxicity (HO-PAA and acetaldehyde) or cytotoxicity (PAA and phenol).

Mots clés: Bacterial metabolites; Oxidative stress; Colonocytes

Mots clés: DNA damage; Cell respiration

Domaines: Sciences du Vivant [q-bio]; Toxicologie

Lucie Armand, Mireille Andriamihaja, Stacy Gellenoncourt, Vincent Bitane, Annaïg Lan, et al.. In vitro impact of amino acid-derived bacterial metabolites on colonocyte mitochondrial activity, oxidative stress response and DNA integrity. Biochimica et Biophysica Acta (BBA) - General Subjects, 2019, 1863 (8), pp.1292-1301. ⟨10.1016/j.bbagen.2019.04.018⟩. ⟨hal-02367934⟩