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

Kun Zhou; Wen Luo; Dan-Dan Gui; Zhong Ren; Dang-Heng Wei; Lu-Shan Liu; Guo-Hua Li; Zhi-Han Tang; Wen-Hao Xiong; Heng-Jing Hu; Zhi-Sheng Jiang

doi:10.1016/j.niox.2023.11.005

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

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

Authors

Kun Zhou¹, Wen Luo², Dan-Dan Gui³, Zhong Ren⁴, Dang-Heng Wei⁵, Lu-Shan Liu⁶, Guo-Hua Li⁷, Zhi-Han Tang⁸, Wen-Hao Xiong⁹, Heng-Jing Hu¹⁰, Zhi-Sheng Jiang¹¹

Affiliations

¹ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 1165797294@qq.com.
² 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. Electronic address: 644584131@qq.com.
³ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 1243756241@qq.com.
⁴ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 35023514@qq.com.
⁵ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 759353094@qq.com.
⁶ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 30186998@qq.com.
⁷ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 453780646@qq.com.
⁸ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: tangzhihan98@163.com.
⁹ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: 52116443@qq.com.
¹⁰ Department of Cardiology, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang City, Hunan Province, 421001, China. Electronic address: bestmanhhj@hotmail.com.
¹¹ Institute of Cardiovascular Disease, Key Lab for Arteriosclerology of Hunan Province, Hengyang Medical School, University of South China, Hengyang, 421001, China. Electronic address: zsjiang2005@163.com.

PMID: 38049061
DOI: 10.1016/j.niox.2023.11.005

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 (H₂S), a protective gaseous mediator in atherosclerosis and the process of EndMT.

Methods: We constructed a stable low-shear-stress-induced(2 dyn/cm²) 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, H₂S 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 H₂S inhibits the binding of NFIL3 and MEST DNA, weakening NFIL3's transcriptional promotion of MEST. Mechanistically, H₂S 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: H₂S negatively regulate the expression of MEST by sulfhydrylation of NFIL3, thereby inhibiting low-shear-stress-induced EndMT and atherosclerosis.

Keywords: Atherosclerosis; EndMT; Hydrogen sulfide; MEST; NFIL3; S-sulfhydrylation.

MeSH terms

Animals
Apolipoproteins E / metabolism
Atherosclerosis* / genetics
Atherosclerosis* / metabolism
DNA / metabolism
Endothelial-Mesenchymal Transition
Endothelium / metabolism
Epithelial-Mesenchymal Transition
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Hydrogen Sulfide* / metabolism
Hydrogen Sulfide* / pharmacology
Mice

Substances

Hydrogen Sulfide
DNA
Apolipoproteins E