C-Reactive protein-induced endothelial microparticle generation in HUVECs is related to BH4-dependent NO formation.
BACKGROUND
C-reactive protein (CRP) has been proven to facilitate endothelial injury via reduced NO production. Endothelial microparticles (EMPs) have emerged as a novel marker of endothelial injury.


METHODS
In vitro cultured human umbilical vein endothelial cells (HUVECs) were incubated with CRP (20 mg/l) for 24 h. The numbers of EMPs with CD31- and CD51-positive staining were assessed flow-cytometrically, and NO production was measured using the Griess reaction in the presence or absence of tetrahydrobiopterin (BH(4)), respectively.


RESULTS
The number of EMPs was significantly increased in HUVECs stimulated by CRP compared with the control group and, in parallel, NO production was decreased (p < 0.05). In the presence of CRP, pretreatment with BH(4) decreased EMP counts and restored NO production to baseline levels (p < 0.05) while pretreatment with 2,4-diamino-6-hydroxypyrimidine (DAHP), a BH(4) synthesis inhibitor, further prompted EMP formation and decreased NO production (p < 0.05). However, adding exogenous BH(4) after pretreatment with DAHP suppressed EMP formation and restored NO production (p < 0.05).


CONCLUSIONS
This study demonstrates that CRP induces EMP generation in HUVECs and this effect is, at least in part, related to impaired BH(4)-dependent NO production. Augmented EMP generation in HUVECs is suggested as a novel potential mechanism contributing to the pathogenesis of vascular injury related to CRP.
Wang
Jie-Mei
JM
Department of Cardiology, First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Wang
Yan
Y
Huang
Jun-Yi
JY
Yang
Zhen
Z
Chen
Long
L
Wang
Li-Chun
LC
Tang
An-Li
AL
Lou
Zhou-Fen
ZF
Tao
Jun
J
D001708:Biopterins / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D002097:C-Reactive Protein / Q000378:metabolism* / Q000494:pharmacology; D048430:Cell Shape; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006136:GTP Cyclohydrolase / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D007042:Hypoxanthines / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D010641:Phenotype; D022161:Transport Vesicles / Q000187:drug effects / Q000378:metabolism*; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D004791:Enzyme Inhibitors; D007042:Hypoxanthines; D001708:Biopterins; D009569:Nitric Oxide; D002097:C-Reactive Protein; D006136:GTP Cyclohydrolase; C017202:2,4-diaminohypoxanthine; C003402:sapropterin