Research Paper Volume 11, Issue 6 pp 1695—1715

Long non-coding RNA CDKN2B-AS1 reduces inflammatory response and promotes cholesterol efflux in atherosclerosis by inhibiting ADAM10 expression

Haocheng Li 1, *, , Song Han 1, *, , Qingfeng Sun 1, , Ye Yao 2, , Shiyong Li 1, , Chao Yuan 1, , Bo Zhang 1, , Bao Jing 1, , Jia Wu 1, , Ye Song 1, , Haiyang Wang 1, ,

  • 1 Department of Vascular Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, P. R. China
  • 2 Department of Cardiac Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, P. R. China
* Equal contribution

received: November 29, 2018 ; accepted: March 6, 2019 ; published: March 29, 2019 ;

https://doi.org/10.18632/aging.101863
How to Cite

Copyright: Li et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Introduction: Long non-coding RNAs (lncRNAs) play key roles in the development of atherosclerosis through the inflammatory pathway. This study aimed to investigate the role of lncRNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) in atherosclerosis via its function in A disintegrin and metalloprotease 10 (ADAM10).

Methods: Initially, the expression of CDKN2B-AS1 and ADAM10 in atherosclerotic plaque tissues and THP-1 macrophage-derived foam cells was determined, after which the cholesterol efflux rate of macrophages was calculated. Interaction between CDKN2B-AS1 and ADAM10 was analyzed, after which, expression of CDKN2B-AS1 and ADAM10 were altered to explore their effects on inflammatory response and cholesterol efflux. The aforementioned findings were further intended to be validated by the atherosclerosis mouse model experiments.

Results: Atherosclerotic plaque tissue and THP-1 macrophage-derived foam cells exhibited downregulated CDKN2B-AS1 and upregulated ADAM10. Upon overexpressing CDKN2B-AS1 or silencing ADAM10, lipid accumulation was reduced and cholesterol efflux was increased. CDKN2B-AS1 located in the nucleus could bind to DNA methyltransferase 1 (DNMT1) to enhance methylation of ADAM10 promoter, leading to suppressed atherosclerotic inflammatory response and promoted cholesterol efflux.

Conclusion: Altogether, lncRNA CDKN2B-AS1 can inhibit the transcription of ADAM10 via DNMT1-mediated ADAM10 DNA methylation, consequently preventing inflammatory response of atherosclerosis and promoting cholesterol efflux.

Abbreviations

LDL: low-density lipoprotein; lncRNAs: long non-coding RNAs; CDKN2B-AS1: cyclin-dependent kinase inhibitor 2B antisense RNA 1; ADAM10: a disintegrin and metalloprotease 10; DNMT1: DNA methyltransferase 1; oe: overexpression; NC: negative control; IMA: internal mammary artery; TNF-ɑ: tumor necrosis factor-ɑ; IL-1β: interleukin-1 β; sh: short hairpin RNA; RIP: RNA-binding protein immunoprecipitation; RPMI: Roswell Park Memorial Institute; FBS: fetal bovine serum; BSA: bovine serum albumin; M.SssI: methyltransferase SssI; RT-qPCR: Reverse transcription-quantitative polymerase chain reaction; UV: ultraviolet; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; PBS: phosphate buffer saline; HPLC: High performance liquid chromatography; BCA: bicinchoninic acid; TC: total cholesterol; FC: free cholesterol; CE: cholesterol ester; ELISA: Enzyme linked immunosorbent assay; RIPA: Radio-Immunoprecipitation Assay; PVDF: polyvinylidene fluoride; TBST: Tris-buffered saline with tween; HRP: horseradish peroxidase; FISH: Fluorescence in situ hybridization; MS-PCR: Methylation-specific PCR; ChIP: Chromatin immunoprecipitation; SPF: specific pathogen-free; TG: triglyceride; HDL: high-density lipoprotein; HE: Hematoxylin-eosin; ANOVA: analysis of variance.