The stretch responsive microRNA miR-148a-3p is a novel repressor of IKBKB, NF-κB signaling, and inflammatory gene expression in human aortic valve cells
The stretch responsive microRNA miR-148a-3p is a novel repressor of IKBKB, NF-κB signaling, and inflammatory gene expression in human aortic valve cells
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DOI：
10.1096/fj.14-257808
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发表时间：
2015-05-01
期刊：
影响因子：
4.8
通讯作者：
Nigam, Vishal
中科院分区：
文献类型：
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作者：
Patel, Vishal;Carrion, Katrina;Nigam, Vishal
二叶式主动脉瓣钙化的速度明显高于正常主动脉瓣，这一过程涉及炎症增加。因为我们以前发现，二叶式主动脉瓣经历更大的拉伸，我们研究了拉伸和人类主动脉瓣间质细胞（AVIC）炎症之间的潜在联系。微阵列，定量PCR（qPCR），和蛋白质检测暴露于周期性拉伸的AVIC显示，拉伸足以增加白细胞介素和金属蛋白酶家族成员的表达超过1.5倍。来自拉伸的AVIC的条件培养基足以激活白细胞。microRNA测序和qPCR实验表明，miR-148a-3p在伸展的AVIC（43%抑制）和作为临床相关的人二叶主动脉瓣（63%减少）中均受到抑制。基于来自qPCR、荧光素酶和Western印迹实验的数据，发现miR-148a-3p是IKBKB的新型阻遏物。此外，增加AVIC中的miR-148 a-3 p水平足以降低NF-κ B（活化B细胞的核因子κ轻链增强子）信号传导和NF-κ B靶基因表达。我们的数据表明，牵张介导的炎症通路活化至少部分是牵张抑制miR-148a-3p和随后抑制IKBKB失败的结果。据我们所知，我们是第一个报道人类AVIC的周期性伸展通过调节中枢炎症通路的microRNA以组织自主方式激活炎症基因的人。
Bicuspid aortic valves calcify at a significantly higher rate than normal aortic valves, a process that involves increased inflammation. Because we have previously found that bicuspid aortic valve experience greater stretch, we investigated the potential connection between stretch and inflammation in human aortic valve interstitial cells (AVICs). Microarray, quantitative PCR (qPCR), and protein assays performed on AVICs exposed to cyclic stretch showed that stretch was sufficient to increase expression of interleukin and metalloproteinase family members by more than 1.5-fold. Conditioned medium from stretched AVICs was sufficient to activate leukocytes. microRNA sequencing and qPCR experiments demonstrated that miR-148a-3p was repressed in both stretched AVICs (43% repression) and, as a clinical correlate, human bicuspid aortic valves (63% reduction). miR-148a-3p was found to be a novel repressor of IKBKB based on data from qPCR, luciferase, and Western blot experiments. Furthermore, increasing miR-148a-3p levels in AVICs was sufficient to decrease NF-kappa B (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling and NF-kappa B target gene expression. Our data demonstrate that stretch-mediated activation of inflammatory pathways is at least partly the result of stretch-repression of miR-148a-3p and a consequent failure to repress IKBKB. To our knowledge, we are the first to report that cyclic stretch of human AVICs activates inflammatory genes in a tissue-autonomous manner via a microRNA that regulates a central inflammatory pathway.