CD39/NTPDase-1 expression and activity in human umbilical vein endothelial cells are differentially regulated by leaf extracts from Rubus caesius and Rubus idaeus

Abstract

Many experimental studies have demonstrated the favorable biological activities of plants belonging to the genus Rubus, but little is known of the role of Rubus leaf extracts in the modulation of the surface membrane expression and activity of endothelial apyrase. The aim of this study was to assess the influence of 1–15 μg/ml Rubus extracts on CD39 expression and enzymatic activity, and on the activation (ICAM-1 expression) and viability of human umbilical vein endothelial cells (HUVEC). The polyphenolic contents and antioxidative capacities of extracts from dewberry (R. caesius L.) and raspberry (R. idaeus L.) leaves were also investigated. The techniques applied were flow cytometry (endothelial surface membrane expression of ICAM-1 and CD39), malachite green assay (CD39 activity), HPLC-DAD (quantitative analysis of polyphenolic extract), ABTS, DPPH and FRAP spectrometric assays (antioxidant capacity), and the MTT test (cell viability). Significantly increased CD39 expressions and significantly decreased ATPDase activities were found in the cells treated with 15 μg/ml of either extract compared to the results for the controls. Neither of the extracts affected cell proliferation, but both significantly augmented endothelial cell ICAM-1 expression. The overall antioxidant capacities of the examined extracts remained relatively high and corresponded well to the determined total polyphenol contents. Overall, the results indicate that under in vitro conditions dewberry and raspberry leaf extracts have unfavorable impact on endothelial cells.

Phenolics and Carotenoid Contents in the Leaves of Different Organic and Conventional Raspberry (Rubus idaeus L.) Cultivars and Their In Vitro Activity

“…One of the most important priorities in research on polyphenolic compound content is not just determining their presence but also biological activity in vitro and in vivo. Dudzińska et al [35] investigated the polyphenol content in raspberry leaves and their antioxidative power. The antioxidant capacities of the examined extracts remained relatively high and corresponded well to the determined total polyphenol content.…”

The influence of Rubus idaeus and Rubus caesius leaf extracts on platelet aggregation in whole blood. Cross-talk of platelets and neutrophils

“…Plant collection and the procedure of extract preparation from raspberry (R. idaeus L.) and dewberry (R. caesius L.) was described previously [1].…”

“…The concentrations of the extracts corresponded to those used in our earlier study of endothelial function (1-15 µg/ml) [1].…”

Tannins as Hemostasis Modulators

“…However, the expression of eNOS in HUVECs was found to be unchanged ( Luzak et al, 2014 ). Similar to the polyphenol fraction of R. nigrum , the tannin-rich polyphenol fraction of the water–acetone extract from R. caesius (10, 15 μg/ml) and R. idaeus (10, 15 μg/ml) leaves increased CD39 expression in HUVECs and decreased its activity (both extracts at a concentration of 15 μg/ml) ( Dudzinska et al, 2014 ).…”

“…Similarly, the procyanidin fraction of an acetone–water extract obtained from Ribes nigrum L. (Grossulariaceae) leaves (10, 30, and 60 μg/ml) inhibited ICAM-1 expression in TNFα-stimulated LT2 endothelial cells ( Garbacki et al, 2005 ). In another study, tannin-rich polyphenol fractions of acetone–water extracts obtained from Rubus caesius L. and Rubus idaeus L. (Rosaceae) leaves increased ICAM-1 expression in unstimulated HUVECs (both fractions at concentrations of 2.5–15 μg/ml) and TNFα-stimulated HUVECs (both fractions at concentrations of 5–15 μg/ml) ( Dudzinska et al, 2014 ). In HUVECs pretreated with LPS, procyanidin B2 (2 µM) decreased inflammasome NLPR3 expression, caspase 1 activation, and interleukin 1β (proinflammatory cytokine) concentration.…”