Identification of side- and shear-dependent microRNAs regulating porcine aortic valve pathogenesis.
Identification of side- and shear-dependent microRNAs regulating porcine aortic valve pathogenesis.
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DOI：
10.1038/srep25397
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发表时间：
2016-05-06
影响因子：
4.6
通讯作者：
Jo H
中科院分区：
文献类型：
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作者：
Rathan S;Ankeny CJ;Arjunon S;Ferdous Z;Kumar S;Fernandez Esmerats J;Heath JM;Nerem RM;Yoganathan AP;Jo H
主动脉瓣（AV）钙化是炎症驱动的过程，优先发生在纤维组织中。为了探索潜在的机制，我们研究了心房中关键的microrna （miRNA）是否由于纤维纤维与脑室相比经历的血流紊乱（振荡剪切（OS））而差异表达。为了鉴定涉及的mirna，从健康猪av的两侧分离内皮富集RNA进行微阵列分析。qPCR验证证实7种mirna （miR-100、-130a、-181a/b、-199a-3p、-199a-5p和-214）在纤维组织中的表达明显高于脑室。经过生物信息学分析，我们选择miR-214在离体剪切系统中使用猪AV小叶进行进一步研究。纤维和脑室侧分别在常规和成骨介质中暴露于振荡或单向脉冲剪切2天，3天和7天。与脑室相比，当纤维瘤暴露于OS时，miR-214的表达更高，纤维瘤的厚度增加，并观察到钙化。anti-miR-214在暴露于OS的整个AV小叶中沉默miR-214显著增加tgf - β1的蛋白表达和适度增加胶原含量，但不影响AV钙化。因此，miR-214被鉴定为侧依赖性和剪切依赖性miRNA，调节AV中的关键机械敏感基因，如tgf - β1。
Aortic valve (AV) calcification is an inflammation driven process that occurs preferentially in the fibrosa. To explore the underlying mechanisms, we investigated if key microRNAs (miRNA) in the AV are differentially expressed due to disturbed blood flow (oscillatory shear (OS)) experienced by the fibrosa compared to the ventricularis. To identify the miRNAs involved, endothelial-enriched RNA was isolated from either side of healthy porcine AVs for microarray analysis. Validation using qPCR confirmed significantly higher expression of 7 miRNAs (miR-100, -130a, -181a/b, -199a-3p, -199a-5p, and -214) in the fibrosa versus the ventricularis. Upon bioinformatics analysis, miR-214 was selected for further investigation using porcine AV leaflets in an ex vivo shear system. Fibrosa and ventricularis sides were exposed to either oscillatory or unidirectional pulsatile shear for 2 days and 3 & 7 days in regular and osteogenic media, respectively. Higher expression of miR-214, increased thickness of the fibrosa, and calcification was observed when the fibrosa was exposed to OS compared to the ventricularis. Silencing of miR-214 by anti-miR-214 in whole AV leaflets with the fibrosa exposed to OS significantly increased the protein expression of TGFβ1 and moderately increased collagen content but did not affect AV calcification. Thus, miR-214 is identified as a side- and shear-dependent miRNA that regulates key mechanosensitive gene in AV such as TGFβ1.