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Updated: Jul 3, 2026
Murine Model of CD40-activation of B cells
Published on: March 5, 2010
Soluble CD40 ligand induces endothelial dysfunction in human and porcine coronary artery endothelial cells
Changyi Chen1, Hong Chai, Xinwen Wang
1Molecular 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
Insights
Soluble CD40 ligand (sCD40L) impairs endothelial function by reducing nitric oxide production and increasing oxidative stress. These effects involve complex molecular pathways impacting eNOS stability, microRNAs, and cellular signaling in endothelial cells.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Endothelial Cell Biology
Background:
- Endothelial dysfunction is a key factor in cardiovascular diseases.
- Soluble CD40 ligand (sCD40L) is implicated in inflammatory processes.
- Understanding sCD40L's impact on endothelial cells is crucial for therapeutic development.
Purpose of the Study:
- To investigate the effects and mechanisms of sCD40L on endothelial dysfunction.
- To analyze sCD40L's impact on endothelial nitric oxide synthase (eNOS) in human coronary artery endothelial cells (HCAECs) and porcine coronary artery rings.
- To elucidate the molecular pathways mediating sCD40L-induced endothelial dysfunction.
Main Methods:
- Treatment of HCAECs and porcine coronary artery rings with sCD40L.
- Measurement of eNOS mRNA and protein levels, mRNA stability, enzyme activity, and nitric oxide (NO) production.
- Assessment of superoxide anion (O(2)(-)) production, mitochondrial membrane potential, and enzyme activities (catalase, SOD, NADPH oxidase).
- Analysis of microRNA expression, MAPK and NF-kappaB signaling pathways.
- Inhibition studies using antioxidants, chemical inhibitors, and dominant-negative mutants.
Main Results:
- sCD40L significantly reduced eNOS mRNA/protein, stability, activity, and NO levels in HCAECs.
- sCD40L increased O(2)(-) production, decreased mitochondrial potential, and altered enzyme activities in HCAECs and porcine arteries.
- sCD40L modulated microRNA expression and activated p38, ERK1/2, and NF-kappaB pathways.
- sCD40L impaired endothelium-dependent vasorelaxation in porcine coronary arteries.
- Specific interventions blocked sCD40L-induced eNOS downregulation.
Conclusions:
- sCD40L induces endothelial dysfunction by decreasing eNOS levels and increasing oxidative stress.
- Molecular mechanisms involve eNOS mRNA stability, microRNAs, mitochondrial dysfunction, and ROS-generating enzymes.
- Key signaling pathways including p38, ERK1/2, and NF-kappaB are activated by sCD40L.
- Targeting these pathways may offer therapeutic strategies for sCD40L-mediated endothelial dysfunction.
Abstract:
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.
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