Effect of acute consumption Passiflora setacea juice on the monocyte adhesion to endothelial cells:a validation ex vivo study

The pulp of Passiflora setacea (PS), a wild species of Brazilian Savanna, has been recently shown to have higherlevels of phenolic compounds, when compared to its commercial species. The leading cause of CVD is atherosclerosis and a diet rich in phenolic compounds may have some effects on monocyte adhesion. Therefore, the aim of the present study was to investigate effect of consumption of PS juice by healthy volunteers on the endothelial cell function, particularly on monocyte adhesion on TNF-alfa-activated endothelial cells.Three middle-aged volunteers were included in a cross-sectional study to analyse the effect of 250mL PS juice consumption. A pull of fasting serum samplings were performed before (t0h) and 3h after the consumption (t3h). Primary human umbilical vein endothelial cells (HUVEC) were cultivated in an endothelial growth medium (EGM) with 25% of volunteers human serum (HS) for 7h with a 4-hour-stimulation of 1 ng/mL of TNF-alfa. Monocytes (THP-1) were added and incubated for 15min with HUVEC. The number of THP1 per HUVEC was counted in microscopic fields.In preliminary results, the treatment with TNF-alfa significantly increased cell adhesion compared with unstimulated control cells (a 3.93-fold increase) (Fig. 2). Pre-exposure of HUVEC to human serum 3 hours after the consumption of PS juice significantly reduced monocyte adherence to TNF-alfa-activated endothelial cells from - 59 to - 43% (P<0,05). The decrease of monocytes adhesion to endothelial cells may indicate an anti-inflammatory and atheroprotective effects of PS juice consumption in healthy volunteer

Anthocyanins and their gut metabolites reduce the adhesion of monocyte to TNFα-activated endothelial cells at physiologically relevant concentrations

An increasing number of evidence suggests a protective role of dietary anthocyanins against cardiovascular diseases. Anthocyanins' extensive metabolism indicates that their metabolites could be responsible for the protective effects associated with consumption of anthocyanin-rich foods. The aim of this work was to investigate the effect of plasma anthocyanins and their metabolites on the adhesion of monocytes to TNFα-activated endothelial cells and on the expression of genes encoding cell adhesion molecules. Human umbilical vein endothelial cells (HUVECs) were exposed to circulating anthocyanins: cyanidin-3-arabinoside, cyanidin-3-galactoside, cyanidin-3-glucoside, delphinidin-3-glucoside, peonidin-3-glucoside, anthocyanin degradation product: 4-hydroxybenzaldehyde, or to their gut metabolites: protocatechuic, vanillic, ferulic and hippuric acid, at physiologically-relevant concentrations (0.1–2 μM) and time of exposure. Both anthocyanins and gut metabolites decreased the adhesion of monocytes to HUVECs, with a magnitude ranging from 18.1% to 47%. The mixture of anthocyanins and that of gut metabolites also reduced monocyte adhesion. However, no significant effect on the expression of genes encoding E-selectin, ICAM1 and VCAM1 was observed, suggesting that other molecular targets are involved in the observed effect. In conclusion, this study showed the potency of anthocyanins and their gut metabolites to modulate the adhesion of monocytes to endothelial cells, the initial step in atherosclerosis development, under physiologically-relevant conditions

Characterization of milk small extracellular vesicles to study adaptation to lactation in ruminants

International audienceSmall extracellular vesicles (EV) are secreted into the extracellular space by all cells. Due to the diversity of their cellular origin and the molecules they contain, small EVs have the ability to ensure extracellular communication and carry molecular signatures of their tissue of origin and its physiological state. In ruminants, early lactation is characterized by profound changes in energy balance and metabolic status. To explore physiological adaptations during early lactation, this study investigated the use of non-invasive milk EVs to identify specific indicators of inter-organ signalling. Milk samples were collected from 8 cows on weeks 2 and 7 postpartum, corresponding to negative and neutral energy balance, respectively. Small EVs were isolated by ultracentrifugation coupled with size exclusion chromatography and characterized by morphological, biophysical and biochemical criteria. Labelled-free shotgun quantitative proteomics was performed by nanoLC-MS/MS. Electron microscopy revealed cup-shaped vesicles with a diameter of about 100 nm, characteristic of small EVs. The diameter was confirmed by Tunable Resistive Pulse Sensing and the specificity of small EVs isolation by the presence of cytoplasmic (TSG101) and membrane (CD63) markers. A total of 508 proteins were identified in milk EVs at weeks 2 and 7 of lactation. Multilevel PCA analysis showed a clear separation between the 2 time points, indicating a strong effect of lactation stage on the protein composition of milk EVs. This proteomic dataset will be analysed to identify small EV molecular signatures of tissue interactions that coordinate nutrient partitioning and adaptation during early lactation