Department of Cardiology, Affiliated Hospital of Gansu Medical College, Kongtong Avenue (East Section), 744000, Pingliang, Gansu Province, China.
Department of Chinese Medicine Management, Affiliated Hospital of Gansu Medical College, 744000, Pingliang, China.
Genes Genomics. 2022 Jun;44(6):671-681. doi: 10.1007/s13258-022-01236-x. Epub 2022 Mar 30.
microRNA-mRNA axes that are involved in oxidized low-density lipoprotein (ox-LDL)-induced vascular smooth muscle cells (VSMCs) proliferation/apoptosis imbalance need to be further investigated.
To investigate the functional role of miR-183-5p/FOXO1 in VSMCs and its interaction with ox-LDL.
RNA sequencing was used to detect transcriptome changes of VSMCs treated with ox-LDL. miR-183-5p and FOXO1 expression levels in VSMCs after ox-LDL treatment were assessed using qRT-PCR and western blotting. The regulatory effect of miR-183-5p on FOXO1 has been tried to prove using a dual-luciferase reporter assay. The functions of miR-183-5p, and FOXO1 were analyzed by CCK-8 assay and flow cytometry assay. The tissue samples or serum samples of high fat-feeding mice and carotid atherosclerosis patients were collected, and the levels of miR-183-5p/FOXO1 were analyzed.
RNA sequencing data showed 81 miRNAs including miR-183-5p was significantly changed after ox-LDL treatment in VSMCs. FOXO1, a miR-183-5p's potential target, was also down-regulated in ox-LDL treated cells. qRT-PCR and western blot found that expression of FOXO1 mRNA and protein significantly reduced in VSMCs treated with ox-LDL, accompanied by overexpression of miR-183-5p. miR-183-5p inhibited FOXO1 mRNA by binding to its 3' UTR. Interference miR-183-5p/FOXO1 could change proliferation/apoptosis imbalance in VSMCs under ox-LDL stimulation. Higher levels of miR-183-5p but reduced FOXO1 can be found in the thoracic aorta tissues of high fat-feeding mice. In serum samples from individuals with carotid atherosclerosis, Higher levels of miR-183-5p were observed. the miR-183-5p level was positively related to the level of serum ox-LDL in patients.
Aberrant expression of miR-183-5p/FOXO1 pathway mediated ox-LDL-induced proliferation/apoptosis imbalance in VSMCs. The miR-183-5p/FOXO1 axis can potentially be utilized as the target in the treatment of patients with atherosclerosis.
涉及氧化型低密度脂蛋白（ox-LDL）诱导的血管平滑肌细胞（VSMCs）增殖/凋亡失衡的 microRNA-mRNA 轴仍需进一步研究。
探讨 miR-183-5p/FOXO1 在 VSMCs 中的功能作用及其与 ox-LDL 的相互作用。
采用 RNA 测序技术检测 ox-LDL 处理后的 VSMCs 转录组变化。采用 qRT-PCR 和 Western blot 检测 ox-LDL 处理后 VSMCs 中 miR-183-5p 和 FOXO1 的表达水平。采用双荧光素酶报告基因实验验证 miR-183-5p 对 FOXO1 的调控作用。通过 CCK-8 检测和流式细胞术检测 miR-183-5p 和 FOXO1 的功能。收集高脂喂养小鼠的组织样本或血清样本和颈动脉粥样硬化患者的血清样本，分析 miR-183-5p/FOXO1 的水平。
RNA 测序数据显示，ox-LDL 处理 VSMCs 后有 81 种 miRNA，包括 miR-183-5p，其表达水平发生显著变化。FOXO1 是 miR-183-5p 的潜在靶基因，在 ox-LDL 处理的细胞中也下调。qRT-PCR 和 Western blot 发现，ox-LDL 处理的 VSMCs 中 FOXO1mRNA 和蛋白表达显著降低，同时 miR-183-5p 过表达。miR-183-5p 通过与其 3'UTR 结合抑制 FOXO1mRNA 的表达。干扰 miR-183-5p/FOXO1 可改变 ox-LDL 刺激下 VSMCs 的增殖/凋亡失衡。高脂喂养小鼠胸主动脉组织中 miR-183-5p 水平升高，FOXO1 水平降低。颈动脉粥样硬化患者血清样本中，miR-183-5p 水平升高。患者血清中 ox-LDL 水平与 miR-183-5p 水平呈正相关。
miR-183-5p/FOXO1 通路的异常表达介导了 ox-LDL 诱导的 VSMCs 增殖/凋亡失衡。miR-183-5p/FOXO1 轴可能成为动脉粥样硬化患者治疗的靶点。
