Combinatory Exposure to Urolithin A, Alternariol, and Deoxynivalenol Affects Colon Cancer Metabolism and Epithelial Barrier Integrity in vitro

Abstract

The human gastrointestinal tract is an important site of nutrient absorption and a crucial barrier against xenobiotics. It regularly faces “chemical cocktails” composed of food constituents, their human and microbial metabolites, and foodborne contaminants, such as mycotoxins. Hence, the colonic epithelium adapts to dietary molecules tuning its immune response, structural integrity, and metabolism to maintain intestinal homeostasis. While gut microbiota metabolites of berry ellagitannins, such as urolithin A (Uro A) might contribute to physiological epithelial barrier integrity, foodborne co-contaminating mycotoxins like alternariol (AOH) and deoxynivalenol (DON) could hamper epithelial function. Hence, we investigated the response of differentiated Caco-2 cells (clone C2BBe1) in vitro to the three compounds alone or in binary mixtures. In virtue of the possible interactions of Uro A, AOH, and DON with the aryl hydrocarbon receptor (AhR) pathway, potential effects on phase-I-metabolism enzymes and epithelial structural integrity were taken as endpoints for the evaluation. Finally, Liquid chromatography tandem mass spectrometry measurements elucidated the absorption, secretion, and metabolic capacity of the cells under single and combinatory exposure scenarios. Uro A and AOH as single compounds, and as a binary mixture, were capable to induce CYP1A1/1A2/1B1 enzymes triggered by the AhR pathway. In light of its ribosome inhibiting capacity, the trichothecene suppressed the effects of both dibenzo-α-pyrones. In turn, cellular responsiveness to Uro A and AOH could be sustained when co-exposed to DON-3-sulfate, instead of DON. Colonic epithelial structural integrity was rather maintained after incubation with Uro A and AOH: this was reinforced in the combinatory exposure scenario and disrupted by DON, an effect, opposed in combination. Passage through the cells as well as the metabolism of Uro A and AOH were rather influenced by co-exposure to DON, than by interaction with each other. Therefore, we conclude that although single foodborne bioactive substances individually could either support or disrupt the epithelial structure and metabolic capacity of colon cancer, exposure to chemical mixtures changes the experimental outcome and calls for the need of combinatory investigations for proper risk assessment.

Effect of the Combined Compound Probiotics with Glycyrrhinic Acid on Alleviating Cytotoxicity of IPEC-J2 Cells Induced by Multi-Mycotoxins

“…This research showed that ADZ addition upregulated the mRNA abundance and downregulated the protein expression of ZO-1. The previous study reported that the cellular mRNA levels were elevated to compensate for the reduction in the protein expression level of ZO-1 [ 59 ], in agreement with this study. Compared with the addition of ADZ alone, CGA addition significantly increased the mRNA abundances of ZO-1 and Claudin-1, but downregulated the protein expression of claudin-1, which was consistent with the previous reports [ 57 , 60 ].…”

Urolithin A induces protective autophagy to alleviate inflammation, oxidative stress, and endoplasmic reticulum stress in pediatric pneumonia

“…20,21 UA has anti-inflammatory and antioxidant properties in vitro and in vivo, and has therapeutic effects in myocardial ischemia-reperfusion injury, brain injury, neuroinflammation, and other diseases. 22 UA inhibits production of hyperglucose-induced neuronal amyloid by regulating ***P < 0.001 vs control; # P < 0.05, ## P < 0.01, ### P < 0.001 vs LPS; @@@ P < 0.001 vs LPS and UA.…”

Microbial Metabolites as Ligands to Xenobiotic Receptors: Chemical Mimicry as Potential Drugs of the Future

“…Another recent advance, at least conceptually, calls for studying microbial metabolites combined at their physiologically relevant concentrations. Groestlinger et al show that combining Urolithin A, Alternariol, and Deoxynivalenol yields different effects on gut barrier function when each is used alone (Groestlinger, Seidl et al 2022). Indeed, newer tryptophan microbial metabolites are being evaluated as AHR ligands and inducers -indole 3carboxaldehyde is an AHR ligand (Vyhlídalová, Krasulová et al 2020) with selective benefit in epithelial cell proliferation rather than intestinal barrier function in piglets (Zhang, Huang et al 2022).…”