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Background  Bovine mammary epithelial cells after calving undergo serious metabolic challenges and oxidative stress both of which could compromise autophagy. Transcription factor EB (TFEB)-mediated autophagy is an important cytoprotective mechanism against oxidative stress. However, effects of TFEB-mediated autophagy on the oxidative stress of bovine mammary epithelial cells remain unknown. Therefore, the main aim of the study was to investigate the role of TFEB-mediated autophagy in bovine mammary epithelial cells experiencing oxidative stress.    Results  H 2 O 2  challenge of the bovine mammary epithelial cell MAC-T increased protein abundance of LC3-II, increased number of autophagosomes and autolysosomes while decreased protein abundance of p62. Inhibition of autophagy via bafilomycin A1 aggravated H 2 O 2 -induced reactive oxygen species (ROS) accumulation and apoptosis in MAC-T cells. Furthermore, H 2 O 2  treatment triggered the translocation of TFEB into the nucleus. Knockdown of TFEB by siRNA reversed the effect of H 2 O 2  on protein abundance of LC3-II and p62 as well as the number of autophagosomes and autolysosomes. Overexpression of TFEB activated autophagy and attenuated H 2 O 2 -induced ROS accumulation. Furthermore, TFEB overexpression attenuated H 2 O 2 -induced apoptosis by downregulating the caspase apoptotic pathway.    Conclusions  Our results indicate that activation of TFEB mediated autophagy alleviates H 2 O 2 -induced oxidative damage by reducing ROS accumulation and inhibiting caspase-dependent apoptosis.

Transcription factor EB (TFEB)-mediated autophagy protects bovine mammary epithelial cells against H2O2-induced oxidative damage in vitro