Inhibition of long noncoding RNA HIF1A-AS2 confers protection against atherosclerosis via ATF2 downregulation

Autor: Di Zhao, Xuemeng Liu, Junhui Xing, Jie-Lei Zhang, Pengcheng Li, Jianwu Jiang, Yanzhou Zhang
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0301 basic medicine
CAD
coronary artery disease
RIP
RNA binding protein immunoprecipitation
lncRNAs
long noncoding RNAs
ICAM-1
intercellular adhesion molecule-1
HCAECs
human coronary artery endothelial cells
USF1
upstream stimulatory factor 1
TNF-α
tumor necrosis factor-α
DMEM
Dulbecco’s modified Eagle’s medium
VCAM-1
vascular cell adhesion molecule 1
0302 clinical medicine
HFD
high fat diet
ChIP
Chromatin immunoprecipitation
LDL
low-density lipoprotein
GAPDH
Glyceraldehyde-3-phosphate dehydrogenase
SMCs
smooth muscle cells
RT-qPCR
reverse transcription quantitative polymerase chain reaction
IL-6
interleukin-6
Gene knockdown
lcsh:R5-920
Multidisciplinary
ND
normal diet
Cell adhesion molecule
Chemistry
sh
short hairpin RNA
MCP-1
monocyte chemoattractant protein-1
030220 oncology & carcinogenesis
HE
Hematoxylin-eosin
Tumor necrosis factor alpha
medicine.symptom
Hypoxia-inducible factor 1 alpha-antisense RNA 2
lcsh:Medicine (General)
ECs
endothelial cells
ATF2
activating transcription factor 2
IL-1β
interleukin-1β
Intercellular Adhesion Molecule-1
IgG
immunoglobulin G
Inflammation
Article
03 medical and health sciences
CCK-8
cell counting kit-8
Activating transcription factor
PBS
phosphate buffered saline
Downregulation and upregulation
ox-LDL
oxidized-low-density lipoprotein
ELISA
enzyme linked immunosorbent assay
medicine
Gene silencing
Upstream transcription factor 1
lcsh:Science (General)
Transcription factor
HIF1A-AS2
hypoxia-inducible factor 1 alpha-antisense RNA 2
ATCC
American Type Culture Collection
Atherosclerosis
si-NC
small interfering RNA-negative control
030104 developmental biology
Cancer research
Long noncoding RNA
lcsh:Q1-390
Zdroj: Journal of Advanced Research, Vol 26, Iss, Pp 123-135 (2020)
Journal of Advanced Research
ISSN: 2090-1232
Popis: Graphical abstract The map illustrating mechanisms associated with lncRNA HIF1A-AS2-mediated inflammation in the atherosclerosis. LncRNA HIF1A-AS2 forms a complex with USF1, which is recruited into the ATF2 promoter to elevate ATF2 expression, thus promoting the development of atherosclerotic inflammation. Meanwhile, downregulation of lncRNA HIF1A-AS2 decreases the expression of ATF2 by reducing the binding of USF1 to the ATF2 promoter regions, thereby inhibiting atherosclerotic inflammation, as reflected by decreased inflammatory factors TNF-α, IL-1β and IL-6 in serum and protein levels of adhesion molecules VCAM-1, ICAM-1, and MCP-1, as well as increased cell viability and reduced apoptosis in ox-LDL-induced inflammation in ECs, SMCs, and HCAECs.
Introduction In atherosclerotic lesions, extensive inflammation of the vessel wall contributes to plaque instability. Long noncoding RNAs (lncRNAs) play important roles in diverse biological processes in atherosclerosis. Objectives Here, we aim to identify the functional role and regulatory mechanisms of lncRNA hypoxia-inducible factor 1 alpha-antisense RNA 2 (HIF1A-AS2) in atherosclerotic inflammation. Methods An atherosclerotic mouse model was induced in ApoE-/- mice by high fat diet (HFD). Endothelial cells (ECs), human aortic smooth muscle cells (SMCs) or human coronary artery endothelial cells (HCAECs) were exposed to ox-LDL to develop the in vitro model. The effects of lncRNA HIF1A-AS2 on inflammation were evaluated by determining levels of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) and levels of adhesion molecules vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and macrophage cationic peptide 1 (MCP-1). Results It was established that lncRNA HIF1A-AS2 and ATF2 were highly expressed in atherosclerotic ApoE-/- mice. Downregulating lncRNA HIF1A-AS2 in ox-LDL-exposed ECs, SMCs and HCAECs inhibited inflammation by reducing levels of pro-inflammatory factors and adhesion molecules. LncRNA HIF1A-AS2 bound to the transcription factor USF1 to elevate ATF2 expression. USF1 overexpression counteracted the suppressive effect of lncRNA HIF1A-AS2 silencing on ox-LDL-induced inflammation. Knockdown of lncRNA HIF1A-AS2 or ATF2 could also attenuate inflammation in atherosclerotic mice. Collectively, the present study demonstrates that downregulation of lncRNA HIF1A-AS2 represses the binding of USF1 to the ATF2 promoter region and then inhibits ATF2 expression, thereby suppressing atherosclerotic inflammation. Conclusion This study suggests lncRNA HIF1A-AS2 as an promising therapeutic target for atherosclerosis.
Databáze: OpenAIRE
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