组蛋白去乙酰化酶 3 通过 microRNA-19b/PPARγ/NF-κB 轴抑制炎症反应来保护动脉粥样硬化。

HDAC3 protects against atherosclerosis through inhibition of inflammation via the microRNA-19b/PPARγ/NF-κB axis.

机构信息

Department of Cardiology, the Second Hospital of Jilin University, Changchun, 130041, PR China.

Department of Radiology, the Second Hospital of Jilin University, Changchun, 130041, PR China.

出版信息

Atherosclerosis. 2021 Apr;323:1-12. doi: 10.1016/j.atherosclerosis.2021.02.013. Epub 2021 Feb 20.

DOI:10.1016/j.atherosclerosis.2021.02.013

PMID:33756273

Abstract

BACKGROUND AND AIMS

Atherosclerosis (AS) is one of the leading causes of cardiovascular diseases. Studies have revealed critical roles of microRNAs (miRNAs) in the progression of AS. This study was conducted to elucidate the role and mechanism by which miR-19b influences AS.

METHODS

Human umbilical vein endothelial cells (HUVECs) were treated with oxidized-low-density lipoprotein (ox-LDL), and an AS mouse model was generated with the help of ApoE mice using a high-fat diet regimen. The expression patterns of peroxisome proliferator-activated receptor γ (PPARγ), nuclear factor κB (NF-κB)/p65, miR-19b and histone deacetylase 3 (HDAC3) were then characterized by reverse transcription quantitative polymerase chain reaction and Western blot analysis. In addition, the relationship among PPARγ, NF-κB/p65, miR-19b and HDAC3 was evaluated by co-immunoprecipitation, chromatin immunoprecipitation and dual-luciferase reporter gene assays. Gain- and loss-of-function experiments were also performed to examine their functional significance on ox-LDL-induced inflammation in HUVECs. Enzyme-linked immunosorbent assay was applied to determine the expression patterns of inflammatory factors in AS mice.

RESULTS

PPARγ and HDAC3 were poorly expressed, while miR-19b and NF-κB/p65 were highly expressed in ox-LDL-induced HUVECs and arterial tissues of AS mice. PPARγ inhibited ox-LDL-induced inflammation in HUVECs by ubiquitination and degradation of NF-κB/p65. miR-19b, downregulated by HDAC3, targeted PPARγ and negatively-regulated its expression. Upregulated PPARγ or HDAC3 or downregulated miR-19b or NF-κB/p65 reduced TNF-α and IL-1β expression levels in ox-LDL-induced HUVECs and AS mice.

CONCLUSIONS

Collectively, the results show that HDAC3 upregulation prevents inflammation to inhibit AS by inactivating NF-κB/p65 via upregulation of miR-19b-mediated PPARγ, providing a basic therapeutic consideration for AS treatment.

摘要

背景与目的

动脉粥样硬化（AS）是心血管疾病的主要病因之一。研究表明 microRNAs（miRNAs）在 AS 的进展中起着关键作用。本研究旨在阐明 miR-19b 影响 AS 的作用机制。

方法

用氧化型低密度脂蛋白（ox-LDL）处理人脐静脉内皮细胞（HUVECs），并用 ApoE 基因敲除小鼠在高脂饮食方案下生成 AS 小鼠模型。然后通过逆转录定量聚合酶链反应和 Western blot 分析来描述过氧化物酶体增殖物激活受体γ（PPARγ）、核因子κB（NF-κB）/p65、miR-19b 和组蛋白去乙酰化酶 3（HDAC3）的表达模式。此外，通过共免疫沉淀、染色质免疫沉淀和双荧光素酶报告基因检测评估了 PPARγ、NF-κB/p65、miR-19b 和 HDAC3 之间的关系。还进行了增益和缺失功能实验，以检查它们对 ox-LDL 诱导的 HUVECs 炎症的功能意义。酶联免疫吸附测定法用于确定 AS 小鼠中炎症因子的表达模式。

结果

在 ox-LDL 诱导的 HUVECs 和 AS 小鼠的动脉组织中，PPARγ 和 HDAC3 的表达水平较低，而 miR-19b 和 NF-κB/p65 的表达水平较高。PPARγ 通过 NF-κB/p65 的泛素化和降解抑制 HUVECs 中的 ox-LDL 诱导的炎症。miR-19b 由 HDAC3 下调，靶向 PPARγ 并负调控其表达。上调 PPARγ 或 HDAC3 或下调 miR-19b 或 NF-κB/p65 可降低 ox-LDL 诱导的 HUVECs 和 AS 小鼠中 TNF-α和 IL-1β的表达水平。