MiR-126 inhibits vascular endothelial cell apoptosis through targeting PI3K/Akt signaling
MiR-126 inhibits vascular endothelial cell apoptosis through targeting PI3K/Akt signaling
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DOI：
10.1007/s00277-015-2567-9
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发表时间：
2016-02-01
影响因子：
3.5
通讯作者：
Du, Kaili
中科院分区：
文献类型：
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作者：
Chen, Lingqiang;Wang, Jing;Du, Kaili
背景：MiR-126 可能与威胁性疾病深静脉血栓 (DVT) 密切相关。目的：本研究旨在探讨异常表达的miR-126对DVT过程中血管内皮细胞（VEC）凋亡的影响，并探讨miR-126在其中的功能。方法：分别用miR-126的antagomir和agomir进行体内miR-126抑制和过表达。采用大鼠创伤性股骨DVT模型，通过TUNEL法和qRT-PCR检测血栓形成前和血栓形成不同时相的VEC凋亡和miR-126表达。通过蛋白质印迹法测量血管组织中 MMP、Akt、Bcl-2、Bad 和 caspase-9 的蛋白水平。在体外，使用 miR-126 抑制剂和模拟物对人脐静脉内皮细胞 (HUVEC) 进行 miR-126 干扰和过表达。用 CoCl2 预处理 HUVEC 后，使用流式细胞术分析细胞凋亡，并使用 qRT-PC/western blotting 测量 miR-126、PIK3R2、PTEN 和磷酸化 Akts 的 RNA/蛋白质水平。结果：抑制 miR-126 可增加 VEC 的凋亡，而过表达 miR-126 可明显挽救 VEC 的凋亡。 PI3K/Akt 信号转导因 miR-126 抑制而受到抑制，并因 miR-126 过表达而明显增强。与这些发现一致的是，PI3K/Akt 通路中的下游蛋白（Bcl-2、Bad 和 cleaved caspase-9）和 MMP 通过抑制或过度表达 miR-126 而发生显着改变。体外实验还表明，当miR-126表达上调时，PI3K/Akt信号传导增强；当miR-126敲低时，PI3K/Akt信号传导受到抑制。结论：过表达的miR-126通过PIK3R2靶向抗凋亡途径PI3K/Akt抑制VEC和DVT的凋亡。一般意义：这些发现可能为 DVT 的治疗提供新的靶点。
Background: MiR-126 is likely to be closely associated with the threatening disease deep venous thrombosis (DVT). Aim: This study aims to investigate the influence of aberrantly expressed miR-126 on vascular endothelial cell (VEC) apoptosis during DVTand explore how miR-126 functions in it. Methods: MiR-126 inhibition and overexpression in vivo were respectively performed with antagomir and agomir of miR-126. Using a rat traumatic femoral DVT model, VEC apoptosis and miR-126 expression were detected by TUNEL assay and qRT-PCR before thrombogenesis and at different time phases of thrombogenesis. Protein levels of MMPs, Akt, Bcl-2, Bad, and caspase-9 in vascular tissue were measured by western blotting. In vitro, miR-126 interference, and overexpression were performed on human umbilical vein endothelial cells (HUVECs) using miR-126 inhibitor and mimics. After HUVECs were pretreated with CoCl2, cell apoptosis was analyzed using flow cytometry, and RNA/protein levels of miR-126, PIK3R2, PTEN, and phosphorylated Akts were measured with qRT-PC/western blotting. Results: The apoptosis of VECs was increased by miR-126 inhibition and obviously rescued by miR-126 overexpression. PI3K/Akt signal transduction was suppressed by miR-126 inhibition and evidently enhanced by miR-126 overexpression. Consistent with these findings, the downstream proteins (Bcl-2, Bad, and cleaved caspase-9) in PI3K/Akt pathway and the MMPs were remarkably changed by inhibition or overexpression of miR-126. In vitro experiments also showed that PI3K/Akt signaling was strengthened when miR-126 expression was upregulated or inhibited when miR-126 was knockdown. Conclusion: Overexpressed miR-126 inhibits apoptosis of VECs and DVT through targeting the anti-apoptotic pathway PI3K/Akt via PIK3R2. General significance: These findings may provide a new target for the therapy of DVT.