AIMS Calcific aortic valve stenosis (CAVS) is the most common valvular heart disease, and is increased with elderly population. However, effective drug therapy has not been established yet. This study aimed to investigate the role of microRNAs (miRs) in the development of CAVS. METHODS AND RESULTS We measured the expression of 10 miRs which were reportedly involved in calcification by using human aortic valve tissue from patients who underwent aortic valve replacement with CAVS or aortic regurgitation (AR) and porcine aortic valve interstitial cells (AVICs) after treatment with osteogenic induction medium. We investigated whether a specific miR-inhibitor can suppress aortic valve calcification in wire injury CAVS mice model. Expression of miR-23a, miR-34a, miR-34c, miR-133a, miR-146a, and miR-155 was increased, and expression of miR-27a and miR-204 was decreased in valve tissues from CAVS compared with those from AR. Expression of Notch1 was decreased, and expression of Runt-related transcription factor 2 (Runx2) was increased in patients with CAVS compared with those with AR. We selected miR-34a among increased miRs in porcine AVICs after osteogenic treatment, which was consistent with results from patients with CAVS. MiR-34a increased calcium deposition in AVICs compared with miR-control. Notch1 expression was decreased, and Runx2 expression was increased in miR-34a transfected AVICs compared with that in miR-control. Conversely, inhibition of miR-34a significantly attenuated these calcification signals in AVICs compared with miR-control. RNA pull down assay revealed that miR-34a directly targeted Notch1 expression by binding to Notch1 mRNA 3' untranslated region. In wire injury CAVS mice, locked nucleic acid miR-34a inhibitor suppressed aortic velocity, calcium deposition of aortic valves, and cardiac hypertrophy, which were involved in decreased Runx2 and increased Notch1 expressions. CONCLUSIONS MiR-34a plays an important role in the development of CAVS via Notch1-Runx2 signaling pathway. Inhibition of miR-34a may be the therapeutic target for CAVS. TRANSLATIONAL PERSPECTIVE This study elucidated that the expression of miR-34a was increased in human specimens of calcific aortic valve stenosis, as well as in vivo and in vitro experiments. MiR-34a plays a role in aortic valve calcification via the Notch1-Runx2 pathway. MiR-34a inhibitor successfully reduced aortic valve calcification in in vivo and in vitro experiments. MiR-34a is a potential therapeutic target for calcific aortic valve stenosis.

中文翻译：

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microRNA-34a的治疗性抑制可通过调节Notch1-Runx2信号传导来改善主动脉瓣钙化。

AIMS钙化主动脉瓣狭窄（CAVS）是最常见的瓣膜性心脏病，并且随着老年人口的增加而增加。但是，尚未建立有效的药物疗法。这项研究旨在调查microRNA（miR）在CAVS的发展中的作用。方法和结果我们使用成骨细胞治疗后用CAVS或主动脉瓣反流（AR）进行主动脉瓣置换术和猪主动脉瓣间质细胞（AVIC）的患者的人主动脉瓣组织测量了据报道参与钙化的10 miRs的表达诱导培养基。我们调查了一种特定的miR抑制剂是否可以抑制导线损伤CAVS小鼠模型中的主动脉瓣钙化。miR-23a，miR-34a，miR-34c，miR-133a，miR-146a和miR-155的表达增加，CAVS的瓣膜组织中的miR-27a和miR-204的表达低于AR。与AR相比，CAVS患者Notch1的表达降低，Runt相关转录因子2（Runx2）的表达增加。我们在成骨治疗后的猪AVICs中增加的miRs中选择了miR-34a，这与CAVS患者的结果一致。与miR对照相比，MiR-34a增加了中航工业中的钙沉积。与miR对照相比，miR-34a转染的AVIC中Notch1表达降低，而Runx2表达增加。相反，与miR对照相比，抑制miR-34a可显着减弱中航工业中的钙化信号。RNA下拉试验显示，miR-34a通过与Notch1 mRNA 3'结合直接靶向Notch1表达 非翻译区域。在钢丝损伤CAVS小鼠中，锁定的核酸miR-34a抑制剂抑制了主动脉速度，主动脉瓣钙沉积和心肌肥大，这与Runx2减少和Notch1表达增加有关。结论MiR-34a通过Notch1-Runx2信号通路在CAVS的发展中起着重要作用。抑制miR-34a可能是CAVS的治疗目标。翻译的角度这项研究阐明了miR-34a在钙化主动脉瓣狭窄的人类标本以及体内和体外实验中的表达增加了。MiR-34a通过Notch1-Runx2途径在主动脉瓣钙化中发挥作用。在体内和体外实验中，MiR-34a抑制剂成功降低了主动脉瓣钙化。