Soluble CD40 ligand induces endothelial dysfunction in human and porcine coronary artery endothelial cells.
The purpose of this study was to determine the effects and mechanisms of sCD40L on endothelial dysfunction in both human coronary artery endothelial cells (HCAECs) and porcine coronary artery rings. HCAECs treated with sCD40L showed significant reductions of endothelial nitric oxide synthase (eNOS) mRNA and protein levels, eNOS mRNA stability, eNOS enzyme activity, and cellular NO levels, whereas superoxide anion (O(2)(-)) production was significantly increased. sCD40L enhanced eNOS mRNA 3'UTR binding to cytoplasmic molecules and induced a unique expression pattern of 95 microRNAs. sCD40L significantly decreased mitochondrial membrane potential, and catalase and SOD activities, whereas it increased NADPH oxidase (NOX) activity. sCD40L increased phosphorylation of MAPKs p38 and ERK1/2 as well as IkappaBalpha and enhanced NF-kappaB nuclear translocation. In porcine coronary arteries, sCD40L significantly decreased endothelium-dependent vasorelaxation and eNOS mRNA levels, whereas it increased O(2)(-) levels. Antioxidant seleno-l-methionine; chemical inhibitors of p38, ERK1/2, and mitochondrial complex II; as well as dominant negative mutant forms of IkappaBalpha and NOX4 effectively blocked sCD40L-induced eNOS down-regulation in HCAECs. Thus, sCD40L reduces eNOS levels, whereas it increases oxidative stress through the unique molecular mechanisms involving eNOS mRNA stability, 3'UTR-binding molecules, microRNAs, mitochondrial function, ROS-related enzymes, p38, ERK1/2, and NF-kappaB signal pathways in endothelial cells.
Chen
Changyi
C
Molecular Surgeon Research Center, Division of Vascular Surgery and Endovascular Therapy, Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, TX, USA. jchen@bcm.tmc.edu
Chai
Hong
H
Wang
Xinwen
X
Jiang
Jun
J
Jamaluddin
Md Saha
MS
Liao
Dan
D
Zhang
Yuqing
Y
Wang
Hao
H
Bharadwaj
Uddalak
U
Zhang
Sheng
S
Li
Min
M
Lin
Peter
P
Yao
Qizhi
Q
D000818:Animals; D000838:Anions / Q000378:metabolism; D023201:CD40 Ligand / Q000378:metabolism*; D003331:Coronary Vessels / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology*; D004730:Endothelium, Vascular / Q000166:cytology*; D005799:Genes, Dominant; D006801:Humans; D020935:MAP Kinase Signaling System; D008954:Models, Biological; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010766:Phosphorylation; D013481:Superoxides / Q000378:metabolism; D013552:Swine
D000838:Anions; D013481:Superoxides; D023201:CD40 Ligand; D052250:Nitric Oxide Synthase Type III; D010100:Oxygen