title: Extract from Ribes nigrum leaves in vitro activates nitric oxide synthase (eNOS) and increases CD39 expression in human endothelial cells.
abstract: The aim of the present study was to evaluate whether blackcurrant leaf extract (BLE) modulates endothelium antithrombotic function, namely increases the expression/activity of ADPase (CD39) and augments the production of nitric oxide in human umbilical vein endothelial cells (HUVEC). It was found that BLE with proanthocyanidins (60 % of the total polyphenol content) increased the CD39-positive endothelial cell fraction (up to 10 % for 2.5 μg/ml, and up to 33 % for 15 μg/ml, p < 0.05 or less) in a concentration-dependent manner, and enhanced endothelial nitric oxide synthase (eNOS) activation (T495 phosphorylation decreased by 31 ± 6 % for 2.5 μg/ml and 48 ± 6 % for 15 μg/ml; S1177 phosphorylation increased by 13 ± 3 % for 2.5 μg/ml and 18 ± 7 % for 15 μg/ml, compared to untreated cells, p < 0.05 or less). Additionally, incubation for 24 or 48 h with BLE at a lower range of polyphenol concentrations, significantly increased cell viability with a maximal effect at 2.5 μg/ml (viability increased by 24.8 ± 1.0 % for 24 h and by 32.5 ± 2.7 % for 48-h time incubation, p < 0.0001). The increased CD39 expression and the increased eNOS activation in HUVEC can be regarded as the beneficial markers of the improvement of antiplatelet action of endothelial cells. Unexpectedly, these assumptions were not confirmed in the experimental model of platelet-endothelial cell interactions. These observations lead to the conclusion that BLE may improve endothelial cell viability at low physiological concentrations without affecting the antiplatelet action of endothelium.
authors.affiliation: Department of Haemostasis and Haemostatic Disorders, Medical University of Lodz, 6/8 Mazowiecka Street, 92-215, Lodz, Poland, boguslawa.luzak@umed.lodz.pl.
authors.forename: Boguslawa
authors.initials: B
authors.lastname: Luzak
authors.forename: Magdalena
authors.initials: M
authors.lastname: Boncler
authors.forename: Joanna
authors.initials: J
authors.lastname: Rywaniak
authors.forename: Dominika
authors.initials: D
authors.lastname: Dudzinska
authors.forename: Marek
authors.initials: M
authors.lastname: Rozalski
authors.forename: Urszula
authors.initials: U
authors.lastname: Krajewska
authors.forename: Ewa
authors.initials: E
authors.lastname: Balcerczak
authors.forename: Anna
authors.initials: A
authors.lastname: Podsedek
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Redzynia
authors.forename: Cezary
authors.initials: C
authors.lastname: Watala
mesh_terms: D015703:Antigens, CD / Q000378:metabolism*; D001081:Apyrase / Q000378:metabolism*; D001792:Blood Platelets / Q000502:physiology; D002450:Cell Communication; D002470:Cell Survival; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D015870:Gene Expression / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism*; D010766:Phosphorylation; D010936:Plant Extracts / Q000494:pharmacology*; D018515:Plant Leaves / Q000737:chemistry; D059808:Polyphenols / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D031965:Ribes / Q000737:chemistry*
chemical_list: D015703:Antigens, CD; D010936:Plant Extracts; D059808:Polyphenols; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D001081:Apyrase; C067869:CD39 antigen