title: 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.
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan; hsatonak-tky@umin.ac.jp.
authors: Hiroshi
authors: H
authors: Satonaka
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Daisuke
authors: D
authors: Nagata
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Masao
authors: M
authors: Takahashi
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Arihiro
authors: A
authors: Kiyosue
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Masahiro
authors: M
authors: Myojo
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Daishi
authors: D
authors: Fujita
authors: Department of Cardiology and Nephrology, Dokkyo Medical University, Kitakobayashi, Mibu, Tochigi, Japan.
authors: Toshihiko
authors: T
authors: Ishimitsu
authors: Graduate School of Pharmaceutical Sciences, University of Tokyo, Hongo, Bunkyo-ku, Tokyo; and.
authors: Tetsuo
authors: T
authors: Nagano
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Ryozo
authors: R
authors: Nagai
authors: Department of Internal Medicine, Graduate School of Medicine, University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan;
authors: Yasunobu
authors: Y
authors: Hirata
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D002448:Cell Adhesion; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism*; D007249:Inflammation / Q000378:metabolism; D008297:Male; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D009131:Muscle, Smooth, Vascular / Q000378:metabolism* / Q000473:pathology; D016328:NF-kappa B / Q000378:metabolism; D058921:Purinergic P2Y Receptor Antagonists / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D058925:Receptors, Purinergic P2Y12 / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: C491766:Ccl2 protein, rat; D018932:Chemokine CCL2; D016328:NF-kappa B; D058921:Purinergic P2Y Receptor Antagonists; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D058925:Receptors, Purinergic P2Y12; D000244:Adenosine Diphosphate
keywords: atherosclerosis; cytokines; inflammation; purinergic P2 receptors; vascular smooth muscle cell