The involvement of the CD40-CD40L pathway in activated platelet-induced changes in HUVEC COX-2 and PPARα expression.
We aim to determine the extent of the CD40-CD40L pathway involvement in activated platelet-induced changes in human umbilical endothelial cells (HUVECs). Activated platelets were co-incubated with HUVECs in the presence or absence of CD40LmAb. HUVECs were also directly stimulated with rhCD40L. HUVEC endothelial cyclooxygenase 2 (COX-2) and peroxisome proliferator-activated receptor alpha (PPARα) expression was then assessed. To estimate COX-2 activity, PGE2 concentration was determined. PPARα activity was assessed using a nuclear factor activity kit. Co-incubation with activated platelets increased HUVEC COX-2 and PPARα mRNA expression (P < 0.01). The addition of CD40L mAb significantly attenuated these increases in mRNA and protein (both P < 0.01). Direct stimulation by rhCD40L increased HUVEC COX-2 mRNA and protein (P < 0.05) but did not significantly change the expression of PPARα mRNA and protein. CD40LmAb significantly decreased (P < 0.05) and rhCD40L significantly (P < 0.01) increased COX-2 enzymatic activity, but had almost no effects on PPARα binding activity. Activated platelets may increase HUVEC COX-2 expression and activity partly through the CD40-CD40L pathway.
Department of Cardiovasology, The Second Affiliated Hospital, Nan Chang University, 1st, Ming-De road, Nanchang, Jiang Xi Province 330006, China. wuyqyq@163.com
Yan-qing
YQ
Wu
Xiao-shu
XS
Chen
Jun-bing
JB
Chai
D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology; D001792:Blood Platelets / Q000502:physiology*; D019013:CD40 Antigens / Q000276:immunology / Q000378:metabolism*; D023201:CD40 Ligand / Q000378:metabolism*; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000096:biosynthesis*; D015232:Dinoprostone / Q000032:analysis; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000276:immunology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D047493:PPAR alpha / Q000096:biosynthesis* / Q000378:metabolism; D015539:Platelet Activation; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055815:Young Adult
D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; D047493:PPAR alpha; D012333:RNA, Messenger; D023201:CD40 Ligand; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D015232:Dinoprostone