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氧化型低密度脂蛋白诱导巨噬细胞凋亡的机制与 microRNA-9 抑制靶基因血小板衍生生长因子受体-β表达的相关性。

Correlation between mechanism of oxidized-low density lipoprotein-induced macrophage apoptosis and inhibition of target gene platelet derived growth factor receptor-β expression by microRNA-9.

机构信息

Department of Cardiology, The Second Hospital of Shanxi Medical University, Taiyuan, China.

Department of Cardiology, Xinzhou People's Hospital, Xinzhou, China.

出版信息

Bioengineered. 2021 Dec;12(2):11716-11725. doi: 10.1080/21655979.2021.2006864.

Abstract

This study was to explore the effects of oxidized-low density lipoprotein (ox-LDL) on the proliferation and apoptosis of macrophages, and the role of miRNA-9 in the targeted regulation of platelet-derived growth factor receptor-β (PDGFR-β) expression. Macrophage RAW264.7 cells were cultured and foamed with 100 mg/L ox-LDL to detect the cell proliferation and apoptosis and target protein expression levels. Subsequently, the miRNA-9 mimics and inhibitors were transfected to detect the expression level of PDGFR-β. The dual-luciferase reporter gene was predicted and applied to detect the target-binding effect of miRNA-9 and PDGFR-β in the cells. The results showed that ox-LDL could induce the foaming of macrophages RAW264.7, inhibit the cell proliferation, and promote the cell apoptosis. After ox-LDL induction, expression of Caspase-3 in macrophages RAW264.7 was up-regulated, and that of glucose regulated protein 78 was down-regulated. The transfection of miRNA-9 mimics could greatly inhibit the expression of PDGFR-β mRNA and proteins in the cells. In addition, the results of the dual-luciferase reporter gene showed that the ratio of luciferase activity was significantly reduced after the miRNA-9 mimic and the wild-type PDGFR-β plasmid were co-transfected. In summary, ox-LDL could induce foaming of macrophages and promote cell apoptosis, and miRNA-9 could target and bind to the 3'UTR region of PDGFR-β, thereby inhibiting the gene expression.

摘要

本研究旨在探讨氧化型低密度脂蛋白(ox-LDL)对巨噬细胞增殖和凋亡的影响,以及 microRNA-9(miRNA-9)在血小板衍生生长因子受体-β(PDGFR-β)表达靶向调控中的作用。培养巨噬细胞 RAW264.7 并以 100mg/L ox-LDL 发泡,检测细胞增殖和凋亡及靶蛋白表达水平。然后转染 miRNA-9 模拟物和抑制剂,检测 PDGFR-β 的表达水平。预测双荧光素酶报告基因并应用于检测细胞中 miRNA-9 与 PDGFR-β 的靶结合效应。结果表明,ox-LDL 可诱导巨噬细胞 RAW264.7 发泡,抑制细胞增殖,促进细胞凋亡。ox-LDL 诱导后,巨噬细胞 RAW264.7 中 Caspase-3 的表达上调,葡萄糖调节蛋白 78 的表达下调。miRNA-9 模拟物的转染可显著抑制细胞中 PDGFR-β mRNA 和蛋白的表达。此外,双荧光素酶报告基因的结果表明,miRNA-9 模拟物与野生型 PDGFR-β 质粒共转染后,荧光素酶活性比值显著降低。综上所述,ox-LDL 可诱导巨噬细胞发泡并促进细胞凋亡,miRNA-9 可靶向结合 PDGFR-β 的 3'UTR 区,从而抑制基因表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b0b4/8810159/c44f6dc0b1d1/KBIE_A_2006864_F0001_B.jpg

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