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EGR-1 decoy ODNs inhibit vascular smooth muscle cell proliferation and neointimal hyperplasia of balloon-injured arteries in rat

机译:EGR-1诱饵ODNs抑制大鼠球囊损伤血管平滑肌细胞增殖和新内膜增生

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Aims: Early growth response factor-1(EGR-1) plays a master regulatory role in multiple cardiovascular pathological processes, such as atherosclerosis and restenosis. For investigating the possibility of using "decoy" strategy to prevent and cure vascular hyperplasia disease, we synthesized the double-stranded, cis-element, decoy oligodeoxynucleotides (ODNs) targeting EGR-1.Main methods: EGR-1 decoy ODNs were transfected into the balloon-injured arteria carotis of rat as well as primary cultures of vascular smooth muscle cells (VSMC). Changes in the thickness of the arterial intima were evaluated by hematoxylin-eosin (HE) staining. VSMC proliferation, DNA synthesis, cell cycle and apoptosis were observed via MTT assay, bromodeoxyuridine (BrdU) incorporation and flow cytometry (FCM). Changes in the expression of EGR-1, and cell cycle related genes, were detected by reverse transcriptase polymerase chain reaction (PT-PCR) and western blot.Key findings: As a result of specific binding to EGR-1 protein, transfected EGR-1 decoy ODNs can reduce EGR-1 promoter affinity, hamper the transcriptional activation of EGR-1-dependent genes, block cell cycle progression of VSMCs, and inhibit neointimal hyperplasia.Significance: Through regulating the cell cycle progression and transcription of target gene, this new "decoy" strategy targeting EGR-1 provides further experimental evidence demonstrating the effectiveness of gene therapy in the treatment of restenosis following percutaneous coronary interventions.
机译:目的:早期生长反应因子-1(EGR-1)在多种心血管病理过程(例如动脉粥样硬化和再狭窄)中起主要调节作用。为了研究使用“诱饵”策略预防和治疗血管增生疾病的可能性,我们合成了靶向EGR-1的双链顺式诱饵寡核苷酸脱氧核苷酸(ODN)。主要方法:将EGR-1诱饵ODN转染到大鼠球囊损伤的动脉以及血管平滑肌细胞(VSMC)的原代培养。通过苏木精-曙红(HE)染色评估动脉内膜厚度的变化。通过MTT测定,溴脱氧尿苷(BrdU)掺入和流式细胞术(FCM)观察到VSMC增殖,DNA合成,细胞周期和凋亡。逆转录酶聚合酶链反应(PT-PCR)和Western blot检测到EGR-1的表达及其与细胞周期相关的基因的变化。 1个诱饵ODNs可以降低EGR-1启动子的亲和力,阻碍EGR-1依赖性基因的转录激活,阻断VSMC的细胞周期进程并抑制新内膜增生。意义:通过调节细胞周期进程和靶基因的转录,针对EGR-1的新“诱饵”策略提供了进一步的实验证据,证明了基因疗法在经皮冠状动脉介入治疗后再狭窄治疗中的有效性。

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