MiR-140-3p is Involved in In-Stent Restenosis by Targeting C-Myb and BCL-2 in Peripheral Artery Disease.
MiR-140-3p is Involved in In-Stent Restenosis by Targeting C-Myb and BCL-2 in Peripheral Artery Disease.
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MiR-140-3p 通过靶向外周动脉疾病中的 C-Myb 和 BCL-2 参与支架内再狭窄
DOI：
10.5551/jat.44024
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发表时间：
2018-11-01
影响因子：
4.4
通讯作者：
Wang SM
中科院分区：
文献类型：
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作者：
Zhu ZR;He Q;Wu WB;Chang GQ;Yao C;Zhao Y;Wang M;Wang SM
目的：支架内再狭窄(ISR)是血管内治疗外周动脉疾病(PAD)后再发缺血和截肢的主要原因。我们先前的研究表明miR-140-3p在PAD动脉中显著下调。然而，miR-140-3p在人PAD的ISR中的表达和功能目前尚不清楚。本研究的目的是确定miR-140-3p在PAD的ISR中的表达及其调节作用。方法：采用实时定量聚合酶链式反应(qRT-PCR)和原位杂交法检测RNA水平。原代培养的ASMC取自健康供者或ISR患者的股动脉。EDU掺入法和CCK-8法检测细胞增殖。Annexin-V/PI双染法和TUNEL法检测细胞凋亡率。采用大鼠颈动脉球囊成形术模型，观察miR-140-3p对再狭窄的影响。结果：与正常动脉相比，PAD和ISR动脉MIR-140-3p表达显著下调。原代培养的ISR ASMCs较正常ASMCs增殖增加，miR-140-3p表达下调。MiR-140-3p可抑制PDGF-BB诱导的ASMCs增殖和诱导细胞凋亡。荧光素酶报告基因分析表明，miR-140-3p通过靶向ISR ASMCs的3‘-UTRs，显著下调ISR ASMCs中C-Myb和bcl2的表达。MIR-140-3p可诱导ASMCs的抗增殖和凋亡作用，而C-Myb或bcl2的过表达可改善上述作用。此外，动物实验表明miR-140-3p可以通过靶向C-Myb和bcl2来减少血管成形术后的再狭窄。结论：miR-140-3p通过靶向C-Myb和bcl-2调控PAD ISR过程中的ASMC功能。这一新发现可能为人类PAD提供一个有希望的治疗靶点。
Aim: In-Stent Restenosis (ISR) is the major reason for recurrent ischemia and amputation after endovascular treatment of Peripheral Artery Disease (PAD). Our previous study demonstrated that miR-140-3p is significantly down-regulated in PAD arteries. However, expression and function of miR-140-3p in ISR of human PAD are currently unclear. The aim of this study is to determine the miR-140-3p expression and its regulative role in ISR of PAD. Methods: The RNA level was determined by quantitative real-time polymerase chain Reaction (qRT-PCR) and in situ hybridization. Primary cultured ASMCs were isolated from human femoral arterial of the healthy donors or ISR patients. Cell proliferation was determined by Edu incorporation and CCK-8 assay. Apoptosis was determined by Annexin-V/PI Double-Staining assay and TUNEL assay. A rat carotid artery balloon angioplasty model was used to investigate the effect of miR-140-3p on restenosis. Results: MiR-140-3p was significantly down-regulated in PAD and ISR arteries than normal arteries. Primary cultured ISR ASMCs exhibited elevated proliferation and down-regulated miR-140-3p than normal ASMCs. Transfection of miR-140-3p mimic attenuated PDGF-BB-induced proliferation in cultured ASMCs and induced apoptosis. Luciferase reporter assay indicated that miR-140-3p transfection significantly down-regulated C-Myb and BCL-2 in ISR ASMCs by targeting to their 3′-UTRs. MiR-140-3p transfection induced anti-proliferation and apoptosis in ASMCs, which were ameliorated by over-expression of C-Myb or BCL-2. Moreover, the animal study showed that miR-140-3p can reduce restenosis following angioplasty via targeting C-Myb and BCL-2. Conclusions: The result suggests that miR-140-3p regulates ASMC function via targeting C-Myb and BCL-2 in the process of ISR in PAD. The novel findings may offer a hopeful therapeutic target for human PAD.