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Nanomolar Caffeic Acid Decreases Glucose Uptake and the Effects of High Glucose in Endothelial Cells.

Natarelli L, Ranaldi G, Leoni G, Roselli M, Guantario B, Comitato R, Ambra R, Cimino F, Speciale A, Virgili F, Canali R - PLoS ONE (2015)

Bottom Line: The decreased flux of glucose associated to caffeic acid affected HG induced apoptosis by down-regulating the expression of initiator (caspase 8 and 9) and effector caspases (caspase 7 and 3) and by increasing the levels of phosphorylated Bcl-2.We also observed that caffeic acid in HG condition was associated to a reduction of p65 subunit nuclear levels with respect to HG alone.In conclusion, our results suggest that caffeic acid, decreasing the metabolic stress induced by HG, allows the activation of survival mechanisms mediated by a different modulation of NF-κB-related signaling pathways and to the activation of anti-apoptotic proteins.

View Article: PubMed Central - PubMed

Affiliation: Council for Agricultural Research and Economics, Food and Nutrition Research Centre, Rome, Italy.

ABSTRACT
Epidemiological studies suggest that moderate and prolonged consumption of coffee is associated with a reduced risk of developing type 2 diabetes but the molecular mechanisms underlying this effect are not known. In this study, we report the effects of physiological concentrations of caffeic acid, easily achievable by normal dietary habits, in endothelial cells cultured in 25 mM of glucose (high glucose, HG). In HG, the presence of 10 nM caffeic acid was associated with a decrease of glucose uptake but not to changes of GLUT-1 membrane localization or mRNA levels. Moreover, caffeic acid countered HG-induced loss of barrier integrity, reducing actin rearrangement and FITC-dextran passage. The decreased flux of glucose associated to caffeic acid affected HG induced apoptosis by down-regulating the expression of initiator (caspase 8 and 9) and effector caspases (caspase 7 and 3) and by increasing the levels of phosphorylated Bcl-2. We also observed that caffeic acid in HG condition was associated to a reduction of p65 subunit nuclear levels with respect to HG alone. NF-κB activation has been shown to lead to apoptosis in HG treated cells and the analysis of the expression of a panel of about 90 genes related to NF-κB signaling pathway revealed that caffeic acid significantly influenced gene expression changes induced by HG. In conclusion, our results suggest that caffeic acid, decreasing the metabolic stress induced by HG, allows the activation of survival mechanisms mediated by a different modulation of NF-κB-related signaling pathways and to the activation of anti-apoptotic proteins.

No MeSH data available.


Related in: MedlinePlus

CA protects HG-dependent loss of endothelial barrier integrity.Cells were seeded onto PET Transwell filters and incubated in control, CA, HG and CA+ HG conditions for 24, 30, 48 and 72 hours. FITC-FD40 was added to a final concentration of 500 μg/ml in the apical medium. The fluorescence was measured both in basal and apical medium. Data are presented as percentage of the ratios of FD40 basal vs apical fluorescence. ***p<0.0001 vs control; n.d. = not detected. Values are the mean ± SEM from at least 3 independent experiments.
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pone.0142421.g002: CA protects HG-dependent loss of endothelial barrier integrity.Cells were seeded onto PET Transwell filters and incubated in control, CA, HG and CA+ HG conditions for 24, 30, 48 and 72 hours. FITC-FD40 was added to a final concentration of 500 μg/ml in the apical medium. The fluorescence was measured both in basal and apical medium. Data are presented as percentage of the ratios of FD40 basal vs apical fluorescence. ***p<0.0001 vs control; n.d. = not detected. Values are the mean ± SEM from at least 3 independent experiments.

Mentions: In order to evaluate the effect of HG on cell-cell contact and barrier integrity, the paracellular diffusion of dextran (40 kDa polymer) was monitored using a fluorescence detector (FITC). At 72 hours of HG incubation, we observed a significant increase of FD40 transit from the apical to the basal medium, indicating the irreversible loss of the barrier integrity because of the generation of membrane perforations. When CA was included in the incubation medium, no changes of the paracellular permeability were observed up to 72 hours of HG treatment (Fig 2).


Nanomolar Caffeic Acid Decreases Glucose Uptake and the Effects of High Glucose in Endothelial Cells.

Natarelli L, Ranaldi G, Leoni G, Roselli M, Guantario B, Comitato R, Ambra R, Cimino F, Speciale A, Virgili F, Canali R - PLoS ONE (2015)

CA protects HG-dependent loss of endothelial barrier integrity.Cells were seeded onto PET Transwell filters and incubated in control, CA, HG and CA+ HG conditions for 24, 30, 48 and 72 hours. FITC-FD40 was added to a final concentration of 500 μg/ml in the apical medium. The fluorescence was measured both in basal and apical medium. Data are presented as percentage of the ratios of FD40 basal vs apical fluorescence. ***p<0.0001 vs control; n.d. = not detected. Values are the mean ± SEM from at least 3 independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4636304&req=5

pone.0142421.g002: CA protects HG-dependent loss of endothelial barrier integrity.Cells were seeded onto PET Transwell filters and incubated in control, CA, HG and CA+ HG conditions for 24, 30, 48 and 72 hours. FITC-FD40 was added to a final concentration of 500 μg/ml in the apical medium. The fluorescence was measured both in basal and apical medium. Data are presented as percentage of the ratios of FD40 basal vs apical fluorescence. ***p<0.0001 vs control; n.d. = not detected. Values are the mean ± SEM from at least 3 independent experiments.
Mentions: In order to evaluate the effect of HG on cell-cell contact and barrier integrity, the paracellular diffusion of dextran (40 kDa polymer) was monitored using a fluorescence detector (FITC). At 72 hours of HG incubation, we observed a significant increase of FD40 transit from the apical to the basal medium, indicating the irreversible loss of the barrier integrity because of the generation of membrane perforations. When CA was included in the incubation medium, no changes of the paracellular permeability were observed up to 72 hours of HG treatment (Fig 2).

Bottom Line: The decreased flux of glucose associated to caffeic acid affected HG induced apoptosis by down-regulating the expression of initiator (caspase 8 and 9) and effector caspases (caspase 7 and 3) and by increasing the levels of phosphorylated Bcl-2.We also observed that caffeic acid in HG condition was associated to a reduction of p65 subunit nuclear levels with respect to HG alone.In conclusion, our results suggest that caffeic acid, decreasing the metabolic stress induced by HG, allows the activation of survival mechanisms mediated by a different modulation of NF-κB-related signaling pathways and to the activation of anti-apoptotic proteins.

View Article: PubMed Central - PubMed

Affiliation: Council for Agricultural Research and Economics, Food and Nutrition Research Centre, Rome, Italy.

ABSTRACT
Epidemiological studies suggest that moderate and prolonged consumption of coffee is associated with a reduced risk of developing type 2 diabetes but the molecular mechanisms underlying this effect are not known. In this study, we report the effects of physiological concentrations of caffeic acid, easily achievable by normal dietary habits, in endothelial cells cultured in 25 mM of glucose (high glucose, HG). In HG, the presence of 10 nM caffeic acid was associated with a decrease of glucose uptake but not to changes of GLUT-1 membrane localization or mRNA levels. Moreover, caffeic acid countered HG-induced loss of barrier integrity, reducing actin rearrangement and FITC-dextran passage. The decreased flux of glucose associated to caffeic acid affected HG induced apoptosis by down-regulating the expression of initiator (caspase 8 and 9) and effector caspases (caspase 7 and 3) and by increasing the levels of phosphorylated Bcl-2. We also observed that caffeic acid in HG condition was associated to a reduction of p65 subunit nuclear levels with respect to HG alone. NF-κB activation has been shown to lead to apoptosis in HG treated cells and the analysis of the expression of a panel of about 90 genes related to NF-κB signaling pathway revealed that caffeic acid significantly influenced gene expression changes induced by HG. In conclusion, our results suggest that caffeic acid, decreasing the metabolic stress induced by HG, allows the activation of survival mechanisms mediated by a different modulation of NF-κB-related signaling pathways and to the activation of anti-apoptotic proteins.

No MeSH data available.


Related in: MedlinePlus