The role of autophagy induction in the mechanism of cytoprotective effect of resveratrol.

We aimed at studying the potential mechanisms in the preventive effect of resveratrol on serum deprivation induced caspase 3 activation on non-transformed cells.

METHODS: Apoptosis was induced by serum deprivation in primary mouse embryonic fibroblasts. Caspase 3 activation, reactive oxygen species production and depolarization of the mitochondrial membrane were measured by fluorescence methods. The involvement of intracellular receptors and autophagy in the effect of resveratrol were analyzed by using specific agonists and antagonists. The role of autophagy was further examined by Western Blot analysis of its protein markers, LC3-II and p62 as well as by acridine orange staining of acidic vacuoles.

RESULTS: We found that neither aromatic hydrocarbon receptors nor estrogen receptors play an important role in the cytoprotective effect of resveratrol. Reactive oxygen species production was not significantly altered by either serum deprivation or resveratrol treatment. In the presence of serum deprivation resveratrol however, induced a significant depolarization in mitochondrial membrane potential. The autophagy inhibitor, chloroquine not only eliminated the preventive effect of resveratrol, but also turned it to deleterious suggesting the prominent role of autophagy induction in the cytoprotective effect. Resveratrol did not alter LC3-II expression, but facilitated p62 degradation in serum deprived cells, suggesting its ability to augment the late phase of autophagy and thus promote the autophagic flux.

CONCLUSION: We have demonstrated that resveratrol can protect primary fibroblasts against serum deprivation induced apoptosis by provoking mild mitochondrial stress and consequent up-regulation of autophagic flux.