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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.

Abbreviations

ABTS:

2,2′-azino-bis(3-ethylbenzo-thiazoline-6 sulfonic acid) radical scavenging activity

ADP:

adenosine 5′-diphosphate sodium salt

ATP:

adenosine 5′-triphosphate disodium salt

ATPDase/NTPDase:

Ca2+/Mg2+-dependent adenosine (ectonucleoside) diphosphohydrolase (apyrase)

BSA:

bovine serum albumin

CE:

catechin equivalents

DMF:

N,N-dimethylformamide

DMSO:

dimethylsulfoxide

DPPH:

2,2-diphenyl-1-picrylhydrazyl radical scavenging activity

EC:

endothelial cells

EDTA:

ethylenediaminetetraacetic acid

FITC:

fluorescein isothiocyanate

FRAP:

ferric reducing antioxidant power assay

GAE:

gallic acid equivalents

HEPES:

4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid

HPLC:

high-performance liquid chromatography

HUVEC:

human umbilical vein endothelial cells

IQR:

interquartile range

LQ:

lower quartile

LSGS:

Low Serum Growth Supplement (Kit)

MTT:

1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazan

PVPP:

polyvinylpolypyrrolidone

R-PE:

Rphycoerythrin

TE:

Trolox equivalents

TNFα:

tumor necrosis factor alpha

TPTZ:

2,4,6-tri-2-pyridyl-s-triazine

Trolox:

6-hydroxy-2,5,7,8-tetramethychroman-2-carboxylic acid

UQ:

upper quartile

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Dudzinska, D., Luzak, B., Boncler, M. et al. CD39/NTPDase-1 expression and activity in human umbilical vein endothelial cells are differentially regulated by leaf extracts from Rubus caesius and Rubus idaeus . Cell Mol Biol Lett 19, 361–380 (2014). https://doi.org/10.2478/s11658-014-0202-8

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Keywords

  • HUVEC
  • Endothelial cells
  • Polyphenolic leaf extracts
  • Rubus
  • Dewberry
  • Raspberry
  • Polyphenols
  • CD39/ATPDase
  • ICAM-1
  • Adhesive molecules
  • Cell activation