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miR-17 通过靶向胰岛素样生长因子 1 调节冠心病内皮细胞的增殖和凋亡。

miR-17 regulates the proliferation and apoptosis of endothelial cells in coronary heart disease via targeting insulin-like-growth factor 1.

机构信息

Department of Cardiology, Zhongda Hospital, Southeast University, Nanjing, 210009, Jiangsu, China.

Department of Cardiology, Zhongda Hospital, Southeast University, Nanjing, 210009, Jiangsu, China.

出版信息

Pathol Res Pract. 2019 Sep;215(9):152512. doi: 10.1016/j.prp.2019.152512. Epub 2019 Jun 20.

Abstract

Coronary heart disease (CHD) is one of the main risks of death, which is mainly caused by coronary arteries arteriosclerosis. The present study aims to investigate the potential roles of miR-17 in CHD. In the present study, Human umbilical vascular endothelial cells (HUVECs) were treated with oxidized low density lipoprotein (ox-LDL). qRT-PCR and western blot were used to examine the mRNA and protein levels, respectively. CCK-8 and flow cytomtry were conducted to determine the proliferation and apoptosis of ox-LDL treated HUVECs. Moreover, luciferase assay was performed to confirm whether insulin-like Growth Factor-1 (IGF-1) was a target of miR-17. The results showed that miR-17 was upregulated in ox-LDL treated HUVECs, while IGF-1 was downregulated. The luciferase activity of ox-LDL treated HUVECs was decreased after the treatment of miR-17 mimics and IGF-1 3'UTR WT. Moreover, overexpressed miR-17 promoted the cell viability and inhibited the apoptosis of ox-LDL treated HUVECs, which was more potent after the treatment of IGF-1 siRNA. Furthermore, the expression of Bax and Caspase3 was decreased, and Bcl-2 was increased in ox-LDL treated HUVECs transfected with miR-17 mimics, which was further decreased after transfection with IGF-1 siRNA. Taken together, miR-17 may regulate the proliferation and apoptosis of ox-LDL treated HUVECs. miR-17 may be a promising biomarker for CHD.

摘要

冠心病(CHD)是主要死亡风险之一,主要由冠状动脉粥样硬化引起。本研究旨在探讨 miR-17 在 CHD 中的潜在作用。在本研究中,用人脐带血管内皮细胞(HUVEC)处理氧化型低密度脂蛋白(ox-LDL)。分别采用 qRT-PCR 和 Western blot 检测 mRNA 和蛋白水平。用 CCK-8 和流式细胞术检测 ox-LDL 处理的 HUVEC 的增殖和凋亡。此外,通过荧光素酶实验证实胰岛素样生长因子-1(IGF-1)是否是 miR-17 的靶基因。结果显示,ox-LDL 处理的 HUVEC 中 miR-17 上调,而 IGF-1 下调。ox-LDL 处理的 HUVEC 的荧光素酶活性在转染 miR-17 模拟物和 IGF-1 3'UTR WT 后降低。此外,过表达 miR-17 促进 ox-LDL 处理的 HUVEC 的细胞活力并抑制其凋亡,在转染 IGF-1 siRNA 后作用更明显。此外,ox-LDL 处理的 HUVEC 中转染 miR-17 模拟物后 Bax 和 Caspase3 的表达降低,Bcl-2 的表达升高,而在转染 IGF-1 siRNA 后进一步降低。总之,miR-17 可能调节 ox-LDL 处理的 HUVEC 的增殖和凋亡。miR-17 可能是 CHD 的有前途的生物标志物。

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