Involvement of P2Y₁₂ receptor in vascular smooth muscle inflammatory changes via MCP-1 upregulation and monocyte adhesion

Abstract

Antiplatelet drugs, frequently used for cardiovascular events with thrombotic involvement, are also regarded as possible promising agents for cardiovascular primary prevention. The roles of P2Y12, an ADP receptor and the target of thienopyridine antiplatelet drugs, are not satisfactorily known in the vascular wall. We investigated the hypothesis that vascular smooth muscle cell (VSMC) P2Y12 is involved in vascular wall inflammatory changes by upregulating monocyte chemoattractant protein-1 (MCP-1) and promoting monocyte adhesion. ADP at 10(-5) M induced a 3.6 ± 0.3-fold upregulation of MCP-1 mRNA in cultured rat VSMCs, which was significantly inhibited by R-138727, the active metabolite of P2Y12 inhibitor prasugrel and siRNAs against P2Y12. ADP also induced MCP-1 protein upregulation, which was diminished by R-138727 and P2Y12 siRNAs. JNK (c-Jun NH2-terminal kinase) inhibition attenuated ADP-induced MCP-1 mRNA and protein upregulation. R-138727 and P2Y12 siRNAs inhibited ADP-induced JNK activation. The reactive oxygen species (ROS) inhibitors N-acetylcysteine (NAC), diphenyleneiodonium (DPI), and Tempol also diminished MCP-1 upregulation and JNK activation induced by ADP. ADP induced MCP-1 promoter activation, which was inhibited by R-138727 and P2Y12 siRNAs. Nuclear factor-κB (NF-κB) consensus sites in the MCP-1 promoter region were involved in this activation. ADP-induced NF-κB pathway activation, examined by a plasmid containing multiple NF-κB sites, was diminished by P2Y12 inhibition. For cellular function analysis, stimulation of VSMC with ADP increased subsequent THP-1 monocyte adhesion. P2Y12 siRNAs and CCR2 antagonism diminished this ADP-induced monocyte adhesion. These data suggested that ADP, via the VSMC P2Y12 receptor, induces vascular inflammatory changes by upregulating MCP-1 and promoting monocyte adhesion.

Myostatin mediates abdominal aortic atherosclerosis progression by inducing vascular smooth muscle cell dysfunction and monocyte recruitment

“…Several in vitro studies performed in cell models revealed the involvement of MAPK signaling pathways in the effects of Mstn3053 and JNK signal has long been recognized to be crucial for negative regulation of muscle growth54. More recently JNK activation in VSMCs has been shown to be essential in ADP-induced MCP-1 expression, promoting monocyte recruitment and vessel wall inflammation14.…”

“…MCP-1 expression, one of the main mediators of vascular inflammation, is triggered by the activation of several transcriptional factors and of different signaling pathways, including MAPkinases, depending on the involved cell type121314.…”

Effects of P2Y12 receptor antagonists beyond platelet inhibition – comparison of ticagrelor with thienopyridines

“…Historically, P2Y 12 receptors have been seen as platelet‐specific. More recently, it has become clear that many cells express P2Y 12 receptors including endothelial cells (EC) (Simon et al, ; Shanker et al, ), vascular smooth muscle cells (VSMC) (Wihlborg et al, ; Satonaka et al, ), dendritic cells (Ben Addi et al, ), leukocytes (Wang et al, ; Diehl et al, ) and neurons (Kawaguchi et al, ). The roles of these non‐platelet P2Y 12 receptors are poorly explored, but data are emerging (Gachet, ).…”

P2Y12 Inhibition beyond Thrombosis: Effects on Inflammation

“…P2Y 12 receptor expression and function in ADP-induced vessel contraction was first described in human internal mammary artery SMCs in 2004 [18]. Activation of P2Y 12 is thought to induce an inflammatory state in VSMCs and correlates with atherosclerotic plaque instability [17,[52][53][54]. It is associated with increased monocyte chemoattractant protein 1 (MCP-1) expression and monocyte adhesion [54].…”

“…Activation of P2Y 12 is thought to induce an inflammatory state in VSMCs and correlates with atherosclerotic plaque instability [17,[52][53][54]. It is associated with increased monocyte chemoattractant protein 1 (MCP-1) expression and monocyte adhesion [54]. P2Y 12 is also implicated in the migration VSMCs through cAMP/PKA signaling pathway associated with actin disassembly and therefore an increase in VSMC motility and migration [55].…”