Chaturvedi Praneet, Chen Neal X, O'Neill Kalisha, McClintick Jeanette N, Moe Sharon M, Janga Sarath Chandra
Department of Biohealth Informatics, School of Informatics and Computing, Indiana University Purdue University, Indianapolis, Indiana, United States of America.
Division of Nephrology, Department of Medicine, School of Medicine, Indiana University, Indianapolis, Indiana, United States of America.
PLoS One. 2015 Jun 26;10(6):e0131589. doi: 10.1371/journal.pone.0131589. eCollection 2015.
Vascular calcification is a complex process and has been associated with aging, diabetes, chronic kidney disease (CKD). Although there have been several studies that examine the role of miRNAs (miRs) in bone osteogenesis, little is known about the role of miRs in vascular calcification and their role in the pathogenesis of vascular abnormalities. Matrix vesicles (MV) are known to play in important role in initiating vascular smooth muscle cell (VSMC) calcification. In the present study, we performed miRNA microarray analysis to identify the dysregulated miRs between MV and VSMC derived from CKD rats to understand the role of post-transcriptional regulatory networks governed by these miRNAs in vascular calcification and to uncover the differential miRNA content of MV. The percentage of miRNA to total RNA was increased in MV compared to VSMC. Comparison of expression profiles of miRNA by microarray demonstrated 33 miRs to be differentially expressed with the majority (~ 57%) of them down-regulated. Target genes controlled by differentially expressed miRNAs were identified utilizing two different complementary computational approaches Miranda and Targetscan to understand the functions and pathways that may be affected due to the production of MV from calcifying VSMC thereby contributing to the regulation of genes by miRs. We found several processes including vascular smooth muscle contraction, response to hypoxia and regulation of muscle cell differentiation to be enriched. Signaling pathways identified included MAP-kinase and wnt signaling that have previously been shown to be important in vascular calcification. In conclusion, our results demonstrate that miRs are concentrated in MV from calcifying VSMC, and that important functions and pathways are affected by the miRs dysregulation between calcifying VSMC and the MV they produce. This suggests that miRs may play a very important regulatory role in vascular calcification in CKD by controlling an extensive network of post-transcriptional targets.
血管钙化是一个复杂的过程，与衰老、糖尿病、慢性肾脏病（CKD）相关。尽管已有多项研究探讨了微小RNA（miRs）在骨成骨中的作用，但对于miRs在血管钙化中的作用及其在血管异常发病机制中的作用知之甚少。已知基质小泡（MV）在启动血管平滑肌细胞（VSMC）钙化中起重要作用。在本研究中，我们进行了miRNA微阵列分析，以鉴定来自CKD大鼠的MV和VSMC之间失调的miRs，以了解这些miRNAs调控的转录后调控网络在血管钙化中的作用，并揭示MV中差异miRNA含量。与VSMC相比，MV中miRNA占总RNA的百分比增加。通过微阵列比较miRNA的表达谱显示33个miRs差异表达，其中大多数（约57%）下调。利用两种不同的互补计算方法Miranda和Targetscan鉴定差异表达miRNAs控制的靶基因，以了解钙化VSMC产生MV可能影响的功能和途径，从而有助于miRs对基因的调控。我们发现包括血管平滑肌收缩、对缺氧的反应和肌肉细胞分化的调控等几个过程显著富集。鉴定出的信号通路包括丝裂原活化蛋白激酶（MAP）-激酶和Wnt信号通路，先前已证明它们在血管钙化中很重要。总之，我们的结果表明，miRs集中在钙化VSMC的MV中，钙化VSMC与其产生的MV之间的miRs失调会影响重要的功能和途径。这表明miRs可能通过控制广泛的转录后靶标网络在CKD的血管钙化中发挥非常重要的调节作用。
