RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification.
RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification.
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对高磷血症刺激的内皮细胞的细胞外囊泡进行 RNA 测序分析，有助于深入了解血管钙化的机制
DOI：
10.1186/s12882-022-02823-6
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发表时间：
2022-05-21
期刊：
影响因子：
2.3
通讯作者：
He, Zhangxiu
中科院分区：
文献类型：
--
作者：
Peng, Zhong;Duan, Yingjie;Zhong, Shuzhu;Chen, Juan;Li, Jianlong;He, Zhangxiu
关键词：
背景高磷血症（HP）与慢性肾脏病（CKD）中的血管钙化（VC）相关。然而，HP诱导的内皮细胞外囊泡（HP-EC-EVs）与VC之间的关系尚不清楚，HP-EC-EVs中miR的表达也尚未确定。方法我们从HP内皮细胞中分离HP-EC-EVs，观察HP-EC-EVs被血管平滑肌细胞（VSMCs）摄取。通过茜素红 S、比色分析和 ALP 活性对 HP-EC-EV 诱导钙沉积进行了表征。为了研究HP-EC-EVs诱导VSMC钙化的机制，对HP-EC-EVs进行了RNA测序。结果我们首先证明HP-EC-EVs在体外诱导VSMC钙化。 HP-EC-EV 的 RNA-seq 分析表明，一种已知的 miR (hsa-miR-3182) 统计上调，12 种 miR 显着下调，这一点通过 qRT-PCR 得到验证。我们通过 miRDB、miRWalk 和 miRanda 数据库分别预测了那些下调和上调 miR 的 58,209 和 74,469 个靶基因。 GO术语显示，下调和上调的靶标主要富集于通过质膜细胞粘附分子（GO：0,016,338，BP）和细胞粘附（GO：0,007,155，BP）、质膜（GO：0,005,886，CC）和金属离子结合的钙依赖性细胞间粘附。 （GO：0,046,914，MF）和 ATP 结合（GO：0,005,524，MF）分别。 KEGG分析的前20个通路包括钙信号通路、cAMP信号通路和ABC转运蛋白，这些通路与VC密切相关。结论我们的结果表明，包装在HP-EC-EV中的那些显着改变的miR可能通过调节相关通路在VC中发挥重要作用。它可能为 CKD 钙化机制提供新的见解。
BackgroundHyperphosphatemia (HP) is associated with vascular calcification (VC) in chronic kidney disease (CKD). However, relationship between HP-induced-endothelial extracellular vesicles (HP-EC-EVs) and VC is unclear, and miR expression in HP-EC-EVs has not been determined.MethodsWe isolated HP-EC-EVs from endothelial cells with HP and observed that HP-EC-EVs were up-taken by vascular smooth muscle cells (VSMCs). HP-EC-EVs inducing calcium deposition was characterized by Alizarin Red S, colourimetric analysis and ALP activity. To investigate the mechanism of HP-EC-EVs-induced VSMC calcification, RNA-sequencing for HP-EC-EVs was performed.ResultsWe first demonstrated that HP-EC-EVs induced VSMC calcification in vitro. RNA-seq analysis of HP-EC-EVs illustrated that one known miR (hsa-miR-3182) was statistically up-regulated and twelve miRs were significantly down-regulated, which was verified by qRT-PCR. We predicted 58,209 and 74,469 target genes for those down- and up-regulated miRs respectively through miRDB, miRWalk and miRanda databases. GO terms showed that down- and up-regulated targets were mostly enriched in calcium-dependent cell–cell adhesion via plama membrane cell-adhesion molecules (GO:0,016,338, BP) and cell adhesion (GO:0,007,155, BP), plasma membrane (GO:0,005,886, CC), and metal ion binding (GO:0,046,914, MF) and ATP binding (GO:0,005,524, MF) respectively. Top-20 pathways by KEGG analysis included calcium signaling pathway, cAMP signaling pathway, and ABC transporters, which were closely related to VC.ConclusionOur results indicated that those significantly altered miRs, which were packaged in HP-EC-EVs, may play an important role in VC by regulating related pathways. It may provide novel insight into the mechanism of CKD calcification.
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影响因子：
3.7
作者：
Eguchi T;Watanabe K;Hara ES;Ono M;Kuboki T;Calderwood SK
通讯作者：
Calderwood SK
DOI：
10.1161/circgenetics.115.001145
发表时间：
2015-12
期刊：
Circulation. Cardiovascular genetics
影响因子：
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作者：
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通讯作者：
Bossé Y
DOI：
10.1159/000450774
发表时间：
2017-01-01
期刊：
SCIENTIFIC ASPECTS OF DIALYSIS THERAPY
影响因子：
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作者：
Massy, Ziad A.;Metzinger-Le Meuth, Valerie;Metzinger, Laurent
通讯作者：
Metzinger, Laurent
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影响因子：
3.7
作者：
Liu J;Xiao X;Shen Y;Chen L;Xu C;Zhao H;Wu Y;Zhang Q;Zhong J;Tang Z;Liu C;Zhao Q;Zheng Y;Cao R;Zu X
通讯作者：
Zu X
影响因子：
2.3
作者：
He, Jin;Zhong, Xiaoyi;Gan, Hua
通讯作者：
Gan, Hua