Diabetes mellitus is generally associated to vascular metabolic dysfunctions and increased endothelial paracellular permeability. Available reports suggest that a moderate and constant coffee consumption is associated with a substantially lower risk of type 2 diabetes. In particular, CAPE (caffeic acid phenetyl ester) has been reported to be able to indirectly modulate the activity of transcription factors involved in stress response, inflammation and detoxification systems. Furthermore, caffeic acid and its metabolites derived from chlorogenic acid, are the most abundant molecules present in circulation after coffee consumption. However, their effects on endothelial cells and their combinatory effects in the presence of high glucose (HG) concentration are not totally known. In this study endothelial cells Ea.hy926, derived from the fusion of HUVEC with tumor A549 cell line were treated with 25 mM of glucose to simulate an hyperglycemic condition and the effect of caffeic acid on high-glucose-dependent increase of permeability and apoptosis was analyzed. Trans-endothelial electrical resistance and the passage of FITC-Dextran were considered markers of the effects of HG on cell permeability in the presence or absence of 10 nM caffeic acid. Morphological changes associated to HG dependent modification of junctional and cytosckeletal protein localization were studied by immunofluorescence techniques and confocal microscopy. The effect of caffeic acid on cell viability in the presence of high glucose was monitored by three independent experiments: trypan blu and total cell count, proliferation analysis with Bromo-Uridin incorporation and citofluorimetric assay of Annexin-V. Our observations indicate that caffeic acid is able to counteract the reduction of electrical resistance associated with HG incubation in a time-dependent manner. On the other hand, the effect of HG and caffeic acid on FITC-dextran permeability was evident only after 72h of treatment. Moreover, caffeic acid countered HG-dependent decrease of cell proliferation and the early apoptosis at 48h from HG treatment. Similarly, caffeic acid totally reversed actin stress fiber formation and cytosolic migration of p120 protein, after 48h of HG treatment. Our results show that physiological concentration of caffeic acid counteracts some of the effects triggered by the exposure to HG on permeability and viability in endothelial cell.
Effect of caffeic acid on endothelial cell dysfunction caused by high-glucose exposure
CIMINO, Francesco;
2012-01-01
Abstract
Diabetes mellitus is generally associated to vascular metabolic dysfunctions and increased endothelial paracellular permeability. Available reports suggest that a moderate and constant coffee consumption is associated with a substantially lower risk of type 2 diabetes. In particular, CAPE (caffeic acid phenetyl ester) has been reported to be able to indirectly modulate the activity of transcription factors involved in stress response, inflammation and detoxification systems. Furthermore, caffeic acid and its metabolites derived from chlorogenic acid, are the most abundant molecules present in circulation after coffee consumption. However, their effects on endothelial cells and their combinatory effects in the presence of high glucose (HG) concentration are not totally known. In this study endothelial cells Ea.hy926, derived from the fusion of HUVEC with tumor A549 cell line were treated with 25 mM of glucose to simulate an hyperglycemic condition and the effect of caffeic acid on high-glucose-dependent increase of permeability and apoptosis was analyzed. Trans-endothelial electrical resistance and the passage of FITC-Dextran were considered markers of the effects of HG on cell permeability in the presence or absence of 10 nM caffeic acid. Morphological changes associated to HG dependent modification of junctional and cytosckeletal protein localization were studied by immunofluorescence techniques and confocal microscopy. The effect of caffeic acid on cell viability in the presence of high glucose was monitored by three independent experiments: trypan blu and total cell count, proliferation analysis with Bromo-Uridin incorporation and citofluorimetric assay of Annexin-V. Our observations indicate that caffeic acid is able to counteract the reduction of electrical resistance associated with HG incubation in a time-dependent manner. On the other hand, the effect of HG and caffeic acid on FITC-dextran permeability was evident only after 72h of treatment. Moreover, caffeic acid countered HG-dependent decrease of cell proliferation and the early apoptosis at 48h from HG treatment. Similarly, caffeic acid totally reversed actin stress fiber formation and cytosolic migration of p120 protein, after 48h of HG treatment. Our results show that physiological concentration of caffeic acid counteracts some of the effects triggered by the exposure to HG on permeability and viability in endothelial cell.Pubblicazioni consigliate
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