Muramyl Dipeptide Induces Reactive Oxygen Species Generation Through the NOD2/COX-2/NOX4 Signaling Pathway in Human Umbilical Vein Endothelial Cells

Abstract: Vascular endothelium dysfunction caused by oxidative stress accelerates the pathologic process of cardiovascular diseases. NOD2, an essential receptor of innate immune system, has been demonstrated to play a critical role in atherosclerosis. Here, the aim of our study was to investigate the effect and underlying molecular mechanism of muramyl dipeptide (MDP) on NOX4-mediated reactive oxygen species (ROS) generation in human umbilical vein endothelial cells (HUVECs). The 2,7-dichlorofluorescein diacetate staining was to measure the intracellular ROS level and showed MDP-promoted ROS production in a time- and dose-dependent manner. The mRNA and protein levels of NOX4 and COX-2 were detected by real-time polymerase chain reaction and western blot. Small interfering RNA (siRNA) was used to silence NOD2 or COX-2 gene expression and investigate the mechanism of NOD2-mediated signaling pathway in HUVECs. Data showed that MDP induced NOX4 and COX-2 expression in a time- and dose-dependent manner. NOD2 knock-down suppressed upregulation of COX-2 and NOX4 in HUVECs treated with MDP. Furthermore, silence of COX-2 in HUVECs downregulated the NOX4 expression after MDP stimulation. Collectively, we indicated that NOD2 played a leading role in MDP-induced COX-2/NOX4/ROS signaling pathway in HUVECs, which was a novel regulatory mechanism in the progress of ROS generation.
Source: Journal of Cardiovascular Pharmacology - Category: Cardiology Tags: Original Article Source Type: research