LncRNA NEAT1 Promote Inflammatory Responses in Coronary Slow Flow Through Regulating miR-148b-3p/ICAM-1 Axis

Qing Zhu; Cuiting Zhao; Yonghuai Wang; Xinxin Li; Yixue Xue; Chunyan Ma

doi:10.2147/JIR.S312583

LncRNA NEAT1 Promote Inflammatory Responses in Coronary Slow Flow Through Regulating miR-148b-3p/ICAM-1 Axis

J Inflamm Res. 2021 Jun 9:14:2445-2463. doi: 10.2147/JIR.S312583. eCollection 2021.

Authors

Qing Zhu¹, Cuiting Zhao¹, Yonghuai Wang¹, Xinxin Li¹, Yixue Xue^{2

3

4}, Chunyan Ma¹

Affiliations

¹ Department of Cardiovascular Ultrasound, The First Hospital of China Medical University, Shenyang, People's Republic of China.
² Department of Neurobiology, School of Life Sciences, China Medical University, Shenyang, People's Republic of China.
³ Key Laboratory of Cell Biology, Ministry of Public Health of China, China Medical University, Shenyang, People's Republic of China.
⁴ Key Laboratory of Medical Cell Biology, Ministry of Education of China, China Medical University, Shenyang, People's Republic of China.

Abstract

Background: Coronary slow flow (CSF) is an angiographic phenomenon characterized by delayed coronary opacification with normal or near-normal epicardial coronary arteries. The pathogenesis of CSF is closely related to inflammatory response. Accumulating evidence shows that long non-coding RNAs (lncRNAs) play an important role in cardiovascular disease. However, the mechanism underlying the influence of the lncRNA nuclear enriched abundant transcripts 1 (NEAT1) on CSF is still unknown.

Patients and methods: Forty CSF patients and forty control subjects were included in the study and underwent coronary angiography, Seattle angina questionnaire (SAQ) and echocardiography. The plasma levels of the inflammatory factors soluble intercellular adhesion molecule-1 (sICAM-1), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) were determined by ELISA. The expression levels of NEAT1, miR-148b-3p and ICAM-1 in cells were measured by qRT-PCR or Western blotting. Cell proliferation was measured by 5-Ethynyl-2'-deoxyuridine (EdU) and Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis was detected by apoptosis assay. The relationship between NEAT1 and miR-148b-3p was verified by luciferase reporter gene assay, RNA immunoprecipitation (RIP) assay and avidin-biotin pull-down assay. The relationship between ICAM-1 and miR-148b-3p was verified by luciferase reporter gene assay and avidin-biotin pull-down assay.

Results: This study showed that plasma sICAM-1, miR-148b-3p, and NEAT1 as independent predictors of a CSF diagnosis. Furthermore, plasma NEAT1 level showed superior diagnostic ability for CSF compared with sICAM-1 and miR-148b-3p. It was also shown that high expression of NEAT1 in oxygen-glucose deprivation (OGD)-treated human umbilical vein endothelial cells (HUVECs) functions as a competing endogenous RNA (ceRNA). By specifically binding miR-148b-3p, it weakened the negative regulatory effects of miR-148b-3p on the ICAM-1 target gene leading to upregulated expression of ICAM-1. This interaction was also shown to inhibit HUVEC proliferation and enhance apoptosis.

Conclusion: This study demonstrated for the first time the important mechanism of action of the NEAT1/miR-148b-3p/ICAM-1 axis in the progression of CSF disease, and indicated the potential of NEAT1, miR-148b-3p, and ICAM-1 as a new target for the diagnosis and treatment of CSF.

Keywords: biomarkers; cell biology; coronary slow flow; inflammation; ncRNA.

Grants and funding

This work was supported by the National Natural Science Foundation of China (grant number 81871373 and 81901766) and the Shenyang Science and Technology Plan Project (grant numbers 20-205-4-014) in study design and in the collection, analysis and interpretation of data.