Reversal of thrombin-induced deactivation of CD39/ATPDase in endothelial cells by HMG-CoA reductase inhibition: effects on Rho-GTPase and adenosine nucleotide metabolism.
Adenosine triphosphate and diphosphate that activate platelet, leukocyte, and endothelium functions are hydrolyzed by endothelial CD39/ATPDase. Because CD39/ATPDase is downregulated in endothelial cells by inflammation and this may be affected by HMG-CoA reductase inhibitors, we examined the role of cerivastatin and simvastatin in regulation of endothelial CD39/ATPDase expression, metabolism of ATP/ADP, and function in platelets. Thrombin-stimulated endothelial cells in vitro were treated with the statins, and hydrolysis of exogenous ADP and ATP was assessed by high-performance liquid chromatography and malachite green assay. Platelet aggregation studies were performed with endothelial cell supernatants as triggers. CD39/ATPDase surface expression by endothelial cells was determined immunologically by fluorescence-activated cell sorter, mRNA expression by RT-PCR, and thrombin-induced dissociation of Rho-GTPases by Western blotting. Treatment by simvastatin or cerivastatin restored impaired metabolism of exogenous ATP and ADP in thrombin-activated endothelial cells by preventing thrombin-induced downregulation of CD39/ATPDase. In platelet aggregation studies, ATP and ADP supernatants of thrombin-activated endothelial cells were less stimulatory in the presence of statins than in their absence. Data show that statins preserve CD39/ATPDase activity in thrombin-treated endothelial cells involving alterations by statins of Rho-GTPase function and CD39/ATPDase expression. Preservation of adenine nucleotide metabolism may directly contribute to the observed anti-thrombotic and anti-inflammatory actions of statins.
Kaneider
Nicole C
NC
Department of Internal Medicine, University of Innsbruck, Austria.
Egger
Petra
P
Dunzendorfer
Stefan
S
Noris
Patrizia
P
Balduini
Carlo L
CL
Gritti
Donatella
D
Ricevuti
Giovanni
G
Wiedermann
Christian J
CJ
D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D015703:Antigens, CD / Q000096:biosynthesis / Q000378:metabolism; D001081:Apyrase / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000378:metabolism*; D001792:Blood Platelets / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology*; D004793:Enzyme Reactivators / Q000493:pharmacokinetics*; D005260:Female; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D007700:Kinetics; D010974:Platelet Aggregation / Q000187:drug effects; D011725:Pyridines / Q000493:pharmacokinetics; D019821:Simvastatin / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology*; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology; D020741:rho GTP-Binding Proteins / Q000378:metabolism
D015703:Antigens, CD; D017077:Culture Media, Conditioned; D004793:Enzyme Reactivators; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D011725:Pyridines; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D019821:Simvastatin; C086276:cerivastatin; D013917:Thrombin; D000251:Adenosine Triphosphatases; D001081:Apyrase; C067869:CD39 antigen; D020741:rho GTP-Binding Proteins