Aflatoxin B1 inhibited autophagy flux by inducing lysosomal alkalinization in HepG2 cells

Aflatoxin B1 (AFB1) is a hazard food pollutant and the most toxic one of all the aflatoxins. It is mainly metabolized in the liver and exerts strong hepatotoxicity and carcinogenesis. Autophagy is an important biological process to maintain the homeostasis of intracellular environment. But the role of autophagy in AFB1-exposured hepatotoxicity remains unclear. The objective of this study was to explore the effect of AFB1 on autophagy flux and its potential mechanisms in HepG2 cells. The data showed AFB1 with no-observed adverse effect level (NOAEL) induced the accumulation of autophagosomes by detecting the level of LC3 and MDC staining. Subsequent findings revealed that autophagosome accumulation was caused by the inhibition of autophagy flux by transfection mRFP-GFP-LC3 adenovirus in the presence of autophagy inhibitor 3-MA and CQ. Further, we investigated lysosomal pH by Acridine orange (AO) and Lysotracker Red (LTR) staining and found that AFB1 exposure caused lysosomal alkalinization. These results indicated AFB1 with NOAEL could inhibit autophagy flux by inducing lysosomal alkalinization. Our study was helpful to further explain early hepatotoxicity mechanism of AFB1.