Warfarin calcifies human aortic valve interstitial cells at high-phosphate conditions via pregnane X receptor
Warfarin calcifies human aortic valve interstitial cells at high-phosphate conditions via pregnane X receptor
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华法林通过孕烷 X 受体在高磷酸盐条件下钙化人主动脉瓣间质细胞
DOI：
10.1007/s00774-019-01001-3
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发表时间：
2019
影响因子：
3.3
通讯作者：
Furukawa Ken-Ichi
中科院分区：
文献类型：
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作者：
Yu Zaiqiang;Seya Kazuhiko;Chiyoya Mari;Daitoku Kazuyuki;Motomura Shigeru;Imaizumi Tadaatsu;Fukuda Ikuo;Furukawa Ken-Ichi
华法林是一种维生素K拮抗剂，是最常见的抗凝剂，用于预防与房颤或瓣膜手术相关的血栓栓塞。虽然一些研究表明，长期使用华法林加速主动脉瓣钙化和主动脉瓣狭窄（AS）的发展，这种现象的详细机制仍不清楚。因此，我们的目的是双重的：建立华法林诱导的钙化的人主动脉瓣间质细胞（HAVIC）使用高无机磷酸盐（Pi）的条件和调查的潜在机制。我们从钙化性主动脉瓣狭窄的主动脉瓣（AS组）和主动脉瓣反流但无任何钙化迹象的主动脉瓣（非AS组）制备并培养HAVIC。在3.2 mM的Pi浓度下，华法林显著增加AS的钙化和碱性磷酸酶（ALP）活性，但不增加非AS组HAVIC。此外，骨形态发生蛋白2（BMP 2）的基因表达，一个钙化标记物，显着增加华法林治疗7天后。在3.2 mM Pi下，Warfarin诱导的AS组HAVICs钙化可被Smad抑制剂dorsomorphin、PXR抑制剂酮康唑和香豆雌酚显著抑制，但不受NF-κB抑制剂SN-50的影响。Warcantine还能够在生理Pi浓度（1.0 mM）下增加BMP 2基因表达。此外，过量的BMP 2（30 ng/mL）促进华法林诱导的ALP上调和HAVIC钙化，在香豆雌酚的存在下，这种作用显著降低。总之，我们的研究结果表明，华法林通过PXR-BMP 2-ALP途径加速AS患者HAVIC的钙化。
Warfarin, a vitamin K antagonist, is the most common anticoagulant used to prevent thromboembolisms associated with atrial fibrillation or following valvular surgery. Although several studies have revealed that long-term warfarin use accelerates aortic valve calcification and the development of aortic stenosis (AS), the detailed mechanism for this phenomenon remains unclear. Therefore, our aim was twofold: to establish the conditions for warfarin-induced calcification of human aortic valve interstitial cells (HAVICs) using high-inorganic phosphate (Pi) conditions and to investigate the underlying mechanism. We prepared and cultured HAVICs from aortic valves affected by calcific aortic valve stenosis (AS group) and aortic valves affected by aortic regurgitation but without any signs of calcification (non-AS group). Under Pi concentrations of 3.2 mM, warfarin significantly increased the calcification and alkaline phosphatase (ALP) activity of AS but not non-AS group HAVICs. Furthermore, gene expression of bone morphogenetic protein 2 (BMP2), a calcigenic marker, was significantly increased following 7 days of warfarin treatment. Warfarin-induced calcification of AS group HAVICs at 3.2 mM Pi was significantly inhibited by dorsomorphin, a Smad inhibitor, and the pregnane X receptor (PXR) inhibitors, ketoconazole and coumestrol, but was unaffected by SN-50, an NF-κB inhibitor. Warfarin was also able to increase BMP2 gene expression at a physiological Pi concentration (1.0 mM). Furthermore, excess BMP2 (30 ng/mL) facilitated warfarin-induced ALP upregulation and HAVIC calcification, an effect which was significantly reduced in the presence of coumestrol. Together, our results suggest that warfarin accelerates calcification of HAVICs from AS patients via the PXR–BMP2–ALP pathway.