Gallic acid protects against endothelial injury by restoring the depletion of DNA methyltransferase 1 and inhibiting proteasome activities

BackgroundThe aim of this study was to investigate the protective effects of gallic acid, a common phenolic compound naturally present in food and nutraceuticals, on endothelial cell death and the mechanisms involved.MethodsEndothelial cell death was induced by the combination of homocysteine, adenosine and tumour necrosis factor (TNF) in human vascular endothelial cells (EAhy926 and HBEC-5i cells). The protective effects of gallic acid were evaluated against cytotoxicity, apoptosis and microparticle release. Underlying mechanisms were further investigated focusing on the involvement of DNA methyltransferase 1 (DNMT1) and proteasome activities.ResultsOur results showed that gallic acid dose-dependently arrested cytotoxicity in EAhy926 and HBEC-5i cells induced by the combination. Gallic acid showed anti-apoptotic effects and reduced the formation of microparticles. Notably, gallic acid reversed DNMT1 depletions at the protein level. The cytoprotective and anti-apoptotic effects of gallic acid were counteracted by the pre-treatment with DNMT1 inhibitor, 5-aza-2-deoxycytidine (5-aza-dC). Treatment with gallic acid led to the accumulation of ubiquitinated protein aggregates and the reduction in chymotrypsin-like proteasome activities indicating proteasome inhibition.ConclusionOur results demonstrate for the first time that gallic acid is capable of protecting endothelial cells from injury induced by the combination of homocysteine, adenosine and TNF, at least in part, by restoring the depletion of DNMT1 and inhibiting proteasome activities.