MicroRNA-30b is a multifunctional regulator of aortic valve interstitial cells
MicroRNA-30b is a multifunctional regulator of aortic valve interstitial cells
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MicroRNA-30b是主动脉瓣间质细胞的多功能调节剂
DOI：
10.1016/j.jtcvs.2013.05.011
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发表时间：
2014-03-01
影响因子：
6
通讯作者：
Xu, Zhiyun
中科院分区：
文献类型：
--
作者：
Zhang, Mi;Liu, Xiaohong;Xu, Zhiyun
目的：主动脉瓣钙化病是一个涉及广泛病理改变的主动过程。瓣膜间质细胞是心脏瓣膜中最常见的细胞，维持着正常的瓣膜结构和功能。MicroRNAs （miRNAs）是基因表达的重要转录后调节剂，miRNA-30b是骨形态发生蛋白2介导的成骨的已知抑制因子。我们假设miRNA-30b是主动脉瓣钙化过程中间质细胞的多功能调节剂。方法：为了确定miRNA-30b在钙化主动脉瓣疾病中的作用，我们评估了术中获得的人钙化主动脉瓣小叶中miRNA的表达。此外，通过定量实时聚合酶链反应、Western blotting、流式细胞术和碱性磷酸酶测定，评估人瓣膜间质细胞miRNA-30b的表达和成骨能力。结果：在本研究中，我们证明了miRNA-30b通过靶向Runx2、Smad1和caspase-3，减弱骨形态发生蛋白2诱导的成骨细胞分化。转染miRNA-30b模拟物导致碱性磷酸酶活性降低，Runx2、Smad1和caspase-3的表达降低。此外，双荧光素酶报告基因检测证实Runx2、Smad1和caspase-3是miRNA-30b的直接靶点。结论：我们通过直接靶向Runx2、Smad1和caspase-3，证明了miRNA-30b作为人主动脉瓣钙化和凋亡的调节剂在钙化性主动脉瓣疾病中的显著作用。靶向miRNA-30b可作为限制主动脉狭窄进行性钙化的新治疗策略。
Objective: Calcific aortic valve disease is an active process involving a wide range of pathologic changes. Valve interstitial cells are the most prevalent cells in the heart valve and maintain normal valve structure and function. MicroRNAs (miRNAs) are essential posttranscriptional modulators of gene expression, and miRNA-30b is a known repressor of bone morphogenetic protein 2-mediated osteogenesis. We hypothesized that miRNA-30b is a multifunctional regulator of aortic valve interstitial cells during calcification.Methods: To determine the role of miRNA-30b in calcific aortic valve disease, we evaluated miRNA expression in human calcific aortic valve leaflets obtained intraoperatively. Furthermore, human valve interstitial cells were evaluated with regard to miRNA-30b expression and osteogenesis by quantitative real-time polymerase chain reaction, Western blotting, flow cytometry, and alkaline phosphatase assays.Results: In this study, we demonstrated that miRNA-30b attenuates bone morphogenetic protein 2-induced osteoblast differentiation by targeting Runx2, Smad1, and caspase-3. Transfection of a mimic of miRNA-30b led to decreases in alkaline phosphatase activity and expressions of Runx2, Smad1, and caspase-3. Furthermore, dual luciferase reporter assays confirmed that Runx2, Smad1, and caspase-3 are direct targets of miRNA-30b.Conclusions: We demonstrated a remarkable role of miRNA-30b in calcific aortic valve disease as a regulator of human aortic valvular calcification and apoptosis through direct targeting of Runx2, Smad1, and caspase-3. Targeting of miRNA-30b could serve as a novel therapeutic strategy to limit progressive calcification in aortic stenosis.