Polyphyenolics increase t-PA and u-PA gene transcription in cultured human endothelial cells.
BACKGROUND
Moderate red wine consumption has been associated with a reduced risk for coronary heart disease, and this cardioprotection may be mediated, in part, by promoting fibrinolysis. This protection may be attributed to the combined or perhaps synergistic effects of alcohol and other red wine components (i.e., polyphenolics). These studies were carried out to determine whether individual phenolics (i.e., catechin, epicatechin, quercetin, and resveratrol) affect fibrinolytic protein (tissue-type plasminogen activator [t-PA] and urokinase-type PA [u-PA]) expression and surface-localized fibrinolytic activity in cultured human umbilical vein endothelial cells (HUVECs).


METHODS
Cultured HUVECs were preincubated (1 hr, 37 degrees C) in the absence or presence of varying concentrations of catechin, epicatechin, quercetin, and resveratrol (0.001-10 microM) and then were washed and incubated for various times in the absence of phenolics. Secreted t-PA/u-PA antigen (24 hr, enzyme-linked immunoadsorbent assay) and mRNA [0-16 hr, reverse transcription-polymerase chain reaction(RT-PCR)] levels and fibrinolytic activity (direct activation of HUVEC-bound 125I-labeled glutamylplasminogen, quantitation of 125I-labeled Mr 20 kDa plasmin light-chain) were measured. Transient transfections of cultured HUVECs were carried out with the pt-PA222/luc and pu-PA236/luc promoter constructs, by using lipofectamine.


RESULTS
Each of the phenolics similarly increased t-PA and u-PA antigen (2- to 3-fold) and mRNA (3- to 4-fold) levels, concomitant with an increase (2- to 3-fold) in sustained (24 hr), surface-localized fibrinolytic activity. Transcription inhibitor actinomycin D abolished the induction of t-PA and u-PA mRNA expression by these phenolics. Transfections with the pt-PA222/luc and pu-PA236/luc promoter constructs showed 2- to 3-fold and 2- to 4-fold increases in luciferase activity for t-PA and u-PA, respectively.


CONCLUSIONS
These results demonstrate that each of these phenolics up-regulates both t-PA and u-PA gene transcription, which results in the sustained increased expression of surface-localized fibrinolytic activity in cultured HUVECs. Wine phenolics increase fibrinolytic activity, independent of ethanol, and it is likely that the overall cardioprotective benefits associated with moderate red wine consumption are attributable to the combined, additive, or perhaps synergistic effects of alcohol and other wine components.
Abou-Agag
L H
LH
Department of Medicine, University of Alabama at Birmingham, 35294-2170, USA.
Aikens
M L
ML
Tabengwa
E M
EM
Benza
R L
RL
Shows
S R
SR
Grenett
H E
HE
Booyse
F M
FM
D002392:Catechin / Q000494:pharmacology; D002478:Cells, Cultured; D003609:Dactinomycin / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism*; D005342:Fibrinolysis; D005419:Flavonoids; D015870:Gene Expression / Q000187:drug effects*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010636:Phenols / Q000494:pharmacology*; D011108:Polymers / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D011794:Quercetin / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D000077185:Resveratrol; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013267:Stilbenes / Q000494:pharmacology; D010959:Tissue Plasminogen Activator / Q000235:genetics*; D014162:Transfection; D014471:Umbilical Veins / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics*
D005419:Flavonoids; D019384:Nucleic Acid Synthesis Inhibitors; D010636:Phenols; D011108:Polymers; D012333:RNA, Messenger; D013267:Stilbenes; D003609:Dactinomycin; D002392:Catechin; D011794:Quercetin; D008156:Luciferases; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D000077185:Resveratrol