Aflatoxin B1 and Aflatoxin M1 Induce Compromised Intestinal Integrity through Clathrin-Mediated Endocytosis

With the growing diversity and complexity of diet, humans are at risk of simultaneous exposure to aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1), which are well-known contaminants in dairy and other agricultural products worldwide. The intestine represents the first barrier against external contaminants; however, evidence about the combined effect of AFB1 and AFM1 on intestinal integrity is lacking. In vivo, the serum biochemical parameters related to intestinal barrier function, ratio of villus height/crypt depth, and distribution pattern of claudin-1 and zonula occluden-1 were significantly affected in mice exposed to 0.3 mg/kg b.w. AFB1 and 3.0 mg/kg b.w. AFM1. In vitro results on differentiated Caco-2 cells showed that individual and combined AFB1 (0.5 and 4 mu g/mL) and AFM1 (0.5 and 4 mu g/mL) decreased cell viability and trans-epithelial electrical resistance values as well as increased paracellular permeability of fluorescein isothiocyanate-dextran in a dose-dependent manner. Furthermore, AFM1 aggravated AFB1-induced compromised intestinal barrier, as demonstrated by the down-regulation of tight junction proteins and their redistribution, particularly internalization. Adding the inhibitor chlorpromazine illustrated that clathrin-mediated endocytosis partially contributed to the compromised intestinal integrity. Synergistic and additive effects were the predominant interactions, suggesting that these toxins are likely to have negative effects on human health.

Automated summary

This study found that exposure to both AFB1 and AFM1 has a significant impact on intestinal integrity, affecting serum biochemical parameters, villus height/crypt depth ratio, and distribution of tight junction proteins in mice. In vitro experiments showed that AFB1 and AFM1 individually and in combination can decrease cell viability and increase paracellular permeability in a dose-dependent manner. Additionally, AFM1 exacerbated the compromised intestinal barrier induced by AFB1, with evidence suggesting that clathrin-mediated endocytosis may play a role in this process. The study also identified synergistic and additive effects between AFB1 and AFM1, indicating a potential negative impact on human health.