Hsa_circ_0004831 的下调部分负责通过靶向 miR-182-5p/CXCL12 轴缓解 ox-LDL 诱导的人脐静脉内皮细胞损伤。

Hsa_circ_0004831 downregulation is partially responsible for atorvastatinalleviated human umbilical vein endothelial cell injuries induced by ox-LDL through targeting the miR-182-5p/CXCL12 axis.

机构信息

Department of Cardiovascular Surgery, The First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Erqi District, Zhengzhou, 453100, China.

Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Erqi District, Zhengzhou, 453100, China.

出版信息

BMC Cardiovasc Disord. 2021 May 1;21(1):221. doi: 10.1186/s12872-021-01998-4.

DOI:10.1186/s12872-021-01998-4

PMID:33932991

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8088699/

Abstract

BACKGROUND

The dysfunction and injury of human umbilical vein endothelial cells (HUVECs) are key events of atherosclerosis (AS). Atorvastatin (ATV) has been shown to play a protective role on endothelial cells. However, the associated molecular mechanisms remain not fully illustrated.

METHODS

HUVECs were treated with oxidized low-density lipoprotein (ox-LDL) to mimic the pathological conditions of endothelial cell injury in AS. Cell injuries were assessed according to cell viability, cell apoptosis, cycle progression, oxidative stress and inflammatory responses using CCK-8 assay, flow cytometry assay or commercial kits. The expression of hsa_circ_0004831, miR-182-5p, and C-X-C motif chemokine 12 (CXCL12) mRNA was examined using quantitative real-time PCR (qPCR). The expression of CXCL12 protein was quantitated by western blot. The predicted target relationship between miR-182-5p and hsa_circ_0004831 or CXCL12 was verified by pull-down assay, dual-luciferase reporter assay or RIP assay.

RESULTS

The expression of hsa_circ_0004831 was upregulated by ox-LDL but downregulated by ATV in HUVECs. ATV promoted cell viability and cell cycle progression but inhibited apoptosis, oxidative stress and inflammation in ox-LDL-treated HUVECs, while the role of ATV was partially reversed by hsa_circ_0004831 overexpression. MiR-182-5p was targeted by hsa_circ_0004831, and hsa_circ_0004831 overexpression-restored apoptosis, oxidative stress and inflammation were blocked by miR-182-5p restoration. Further, CXCL12 was targeted by miR-182-5p, and miR-182-5p inhibition-stimulated apoptosis, oxidative stress and inflammation were lessened by CXCL12 knockdown.

CONCLUSION

Hsa_circ_0004831-targeted miR-182-5p/CXCL12 regulatory network is one of the pathways by which ATV protects against ox-LDL-induced endothelial injuries.

摘要

背景

人脐静脉内皮细胞（HUVEC）功能障碍和损伤是动脉粥样硬化（AS）的关键事件。阿托伐他汀（ATV）已被证明对内皮细胞具有保护作用。然而，相关的分子机制仍未完全阐明。

方法

用氧化型低密度脂蛋白（ox-LDL）处理 HUVEC，模拟 AS 内皮细胞损伤的病理状态。根据 CCK-8 测定、流式细胞术测定或商业试剂盒，用细胞活力、细胞凋亡、周期进程、氧化应激和炎症反应评估细胞损伤。采用实时定量 PCR（qPCR）检测 hsa_circ_0004831、miR-182-5p 和 C-X-C 基序趋化因子 12（CXCL12）mRNA 的表达。用 Western blot 定量 CXCL12 蛋白的表达。通过下拉测定、双荧光素酶报告测定或 RIP 测定验证 miR-182-5p 与 hsa_circ_0004831 或 CXCL12 的预测靶关系。

结果

ox-LDL 上调 HUVEC 中 hsa_circ_0004831 的表达，而 ATV 下调其表达。ATV 促进 ox-LDL 处理的 HUVEC 中细胞活力和细胞周期进程，但抑制细胞凋亡、氧化应激和炎症，而过表达 hsa_circ_0004831 部分逆转了 ATV 的作用。miR-182-5p 是 hsa_circ_0004831 的靶标，hsa_circ_0004831 过表达恢复的凋亡、氧化应激和炎症被 miR-182-5p 恢复所阻断。此外，CXCL12 是 miR-182-5p 的靶标，miR-182-5p 抑制刺激的凋亡、氧化应激和炎症在 CXCL12 敲低时减轻。