硫化氢通过巯基化内皮 NFIL3 抑制 MEST 介导的内皮间质转化来减轻低切应力诱导的动脉粥样硬化。

Hydrogen sulfide attenuates atherosclerosis induced by low shear stress by sulfhydrylating endothelium NFIL3 to restrain MEST mediated endothelial mesenchymal transformation.

机构信息

Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China.

Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China; Department of Basic Medicine, Changsha Health Vocational College, Changsha, 410699, China.

出版信息

Nitric Oxide. 2024 Jan 1;142:47-57. doi: 10.1016/j.niox.2023.11.005. Epub 2023 Dec 2.

DOI:10.1016/j.niox.2023.11.005

PMID:38049061

Abstract

BACKGROUND

Endothelial-mesenchymal transition (EndMT) induced by low shear stress plays an important role in the development of atherosclerosis. However, little is known about the correlation between hydrogen sulfide (HS), a protective gaseous mediator in atherosclerosis and the process of EndMT.

METHODS

We constructed a stable low-shear-stress-induced(2 dyn/cm) EndMT model, acombined with the pretreatment method of hydrogen sulfide slow release agent(GYY4137). The level of MEST was detected in the common carotid artery of ApoE mice with local carotid artery ligation. The effect of MEST on atherosclerosis development in vivo was verified using ApoE mice were given tail-vein injection of endothelial-specific overexpressed and knock-down MEST adeno-associated virus (AAV).

RESULTS

These findings confirmed that MEST is up-regulated in low-shear-stress-induced EndMT and atherosclerosis. In vivo experiments showed that MEST gene overexpression significantly promoted EndMT and aggravated the development of atherosclerotic plaques and MEST gene knockdown significantly inhibited EndMT and delayed the process of atherosclerosis. In vitro, HS inhibits the expression of MEST and EndMT induced by low shear stress and inhibits EndMT induced by MEST overexpression. Knockdown of NFIL3 inhibit the up regulation of MEST and EndMT induced by low shear stress in HUVECs. CHIP-qPCR assay and Luciferase Reporter assay confirmed that NFIL3 binds to MEST DNA, increases its transcription and HS inhibits the binding of NFIL3 and MEST DNA, weakening NFIL3's transcriptional promotion of MEST. Mechanistically, HS increased the sulfhydrylation level of NFIL3, an important upstream transcription factors of MEST. In part, transcription factor NFIL3 restrain its binding to MEST DNA by sulfhydration.

CONCLUSIONS

HS negatively regulate the expression of MEST by sulfhydrylation of NFIL3, thereby inhibiting low-shear-stress-induced EndMT and atherosclerosis.

摘要

背景

低切应力诱导的内皮-间充质转化（EndMT）在动脉粥样硬化的发生发展中起着重要作用。然而，关于在动脉粥样硬化中起保护作用的气态介质硫化氢（H2S）与EndMT 过程之间的相关性知之甚少。

方法

我们构建了一个稳定的低切应力诱导（2 dyn/cm）EndMT 模型，并结合了硫化氢缓慢释放剂（GYY4137）的预处理方法。通过局部颈动脉结扎，检测载脂蛋白 E（ApoE）小鼠颈总动脉中 MEST 的水平。通过尾静脉注射内皮特异性过表达和敲低 MEST 的腺相关病毒（AAV），在 ApoE 小鼠体内验证 MEST 对动脉粥样硬化发展的影响。

结果

这些发现证实 MEST 在低切应力诱导的 EndMT 和动脉粥样硬化中上调。体内实验表明，MEST 基因过表达显著促进 EndMT 并加重动脉粥样硬化斑块的发展，而 MEST 基因敲低则显著抑制 EndMT 并延缓动脉粥样硬化进程。在体外，H2S 抑制低切应力诱导的 MEST 表达和 EndMT，并抑制 MEST 过表达诱导的 EndMT。NFIL3 敲低抑制 HUVECs 中低切应力诱导的 MEST 和 EndMT 的上调。CHIP-qPCR 分析和荧光素酶报告基因检测证实，NFIL3 与 MEST DNA 结合，增加其转录，而 H2S 抑制 NFIL3 与 MEST DNA 的结合，减弱 NFIL3 对 MEST 的转录促进作用。机制上，H2S 增加了 MEST 的重要上游转录因子 NFIL3 的巯基化水平。在某种程度上，转录因子 NFIL3 通过巯基化来抑制其与 MEST DNA 的结合。