Tohoku J. Exp. Med., 2019 December, 249(4)
Adenosine Attenuates Aortic Smooth Muscle Cell Calcification through A3 Adenosine Receptor
KENTA FUJIMOTO,1 ATSUSHI SHIOI,2,3 YUYA MIKI,1 YOSHINORI KAKUTANI,1 TOMOAKI MORIOKA,1 TETSUO SHOJI,2,3 MASANORI EMOTO1 and MASAAKI INABA1,3
1Department of Metabolism, Endocrinology, and Molecular Medicine, Osaka City University Graduate School of Medicine, Osaka, Osaka, Japan
2Department of Vascular Medicine, Osaka City University Graduate School of Medicine, Osaka, Osaka, Japan
3Vascular Science Center for Translational Research, Osaka City University Graduate School of Medicine, Osaka, Osaka, Japan
Vascular calcification is a typical feature of atherosclerosis and is associated with adverse cardiovascular events such as myocardial infarction and stroke. Several studies have suggested that adenosine, an ATP metabolite may function as an endogenous regulator of arterial calcification. However, its effects on vascular smooth muscle cell calcification have not been clarified. In this study, we investigated the inhibitory effects of adenosine on vascular calcification in vitro by utilizing the culture of human aortic smooth muscle cells (HASMCs). Osteoblastic differentiation of HASMCs was induced by the treatment with oncostatin M and osteogenic differentiation medium. Adenosine and its metabolically stable analogue, 2-chloroadenosine (CADO) significantly reduced matrix mineralization and alkaline phosphatase (ALP) activities in HASMCs. The mRNA expression of tissue non-specific alkaline phosphatase (TNAP) was down-regulated by adenosine and CADO, but the mRNA expression of other osteoblastic differentiation markers, such as Runt-related transcription factor 2 (RUNX2) and bone sialoprotein (BSP)-II, was not significantly affected by these two reagents. Among the adenosine receptor (AR) subtype-selective agonists used, only IB-MECA (A3 AR-selective agonist) significantly decreased in vitro mineralization and ALP activities in HASMCs, but not with CCPA (A1 AR-selective agonist), CGS21680 (A2a AR-selective agonist), or BAY60-6583 (A2b AR-selective agonist). Importantly, IB-MECA also down-regulated expression of TNAP mRNA. Finally, knockdown of A3 AR, but not A1 AR, A2a AR, or A2b AR, significantly reversed the inhibitory actions of adenosine, CADO, or IB-MECA on in vitro calcification and ALP activities in HASMCs. These data suggest that adenosine attenuates HASMC calcification through A3 AR.
Keywords —— A3 adenosine receptor; adenosine; oncostatin M; osteoblastic differentiation; vascular smooth muscle cells
© 2019 Tohoku University Medical Press
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Tohoku J. Exp. Med., 2019, 249, 275-283
Correspondence: Atsushi Shioi, M.D., Ph.D., Department of Vascular Medicine, Osaka City University Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka, Osaka 545-8585, Japan.
e-mail: as@med.osaka-cu.ac.jp