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Circular RNA circ_0001445 alleviates the ox-LDL-induced endothelial injury in human primary aortic endothelial cells through regulating ABCG1 via acting as a sponge of miR-208b-5p

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Abstract

Background

Coronary artery disease (CAD) originates from the blockage of the inner walls of the coronary arteries due to a plaque buildup. Circular RNA (circRNA) circ_0001445 has been reported to be downregulated in patients with a higher coronary atherosclerotic burden. This study is designed to explore the role and mechanism of circ_0001445 on the oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell damage.

Methods

Circ_0001445, microRNA-208b-5p (miR-208b-5p), and ATP-binding cassette sub-family G member 1 (ABCG1) levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Inflammatory cytokines levels, cell viability, proliferation, migration were detected by Enzyme-linked immunosorbent assay (ELISA) kits, Cell Counting Kit-8 (CCK-8), 5-ethynyl-2’-deoxyuridine (EdU), and transwell assays, respectively. Protein levels were determined by western blot assay. The binding between miR-208b-5p and circ_0001445 or ABCG1 was predicted by circBank or TargetScan, and then verified by a dual-luciferase reporter, RNA Immunoprecipitation (RIP), and RNA pull-down assays.

Results

Circ_0001445 and ABCG1 were decreased, and miR-208b-5p was increased in CAD patients and ox-LDL-treated HAECs. Also, circ_0001445 overexpression could weaken ox-LDL-triggered HAEC injury by boosting proliferation, migration, and repressing inflammation and extracellular matrix (ECM). Mechanically, circ_0001445 directly targeted miR-208b-5p. Furthermore, miR-208b-5p mediated the modulation of circ_0001445 in ox-LDL-induced HAEC injury. ABCG1 acted as a direct target of miR-208b-5p, and the downregulation of miR-208b-5p relieved ox-LDL-induced HAEC damage by interacting with ABCG1. Additionally, circ_0001445 regulated ABCG1 expression by sponging miR-208b-5p.

Conclusion

Circ_0001445 could abate ox-LDL-mediated HAEC damage by the miR-208b-5p/ABCG1 axis, providing a novel insight into the pathogenesis and treatment of CAD.

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Data availability

The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.

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Authors and Affiliations

  1. Department of Cardiovascular Medicine, 920 Hospital of Joint Logistics Support Force, No. 212, Daguan Road, Xishan District, Kunming, Yunnan Province, China

    Zhihua Yang & Lixia Yang

  2. Department of Cardiology, The Second Affiliated Hospital of Chongqing Medical University, 288 Tianwen Avenue, Nan’an District, Chongqing, China

    Xing Liang

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Correspondence to Lixia Yang.

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The present study was approved by the ethical review committee of 920 Hospital of Joint Logistics Support Force. Written informed consent was obtained from all enrolled patients.

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Supplementary file1 (TIF 270 KB)

11748_2022_1799_MOESM2_ESM.tif

Supplementary file2 (TIF 1504 KB) Figure S1 circ_0001445 expression was reduced in CAD patients. (A) RT-qPCR analysis of circ_0001445 expression in serum samples from 26 healthy volunteers and 44 CAD patients (24 Angina pectoris, 6 Myocardial infarction, 9 Asymptomatic myocardial ischemia, 7 Ischemic cardiomyopathy, 4 Sudden death coronary heart disease). (B) ROC curve analysis for the diagnostic value evaluation of circ_0001445 in CAD patients. **P <0.01, ***P <0.001. Figure S2 The effects of circ_0001445 knockdown ox-LDL-mediated inflammatory response, proliferation, migration, and ECM in HAECs. (A) RT-qPCR assay was used to detect circ_0001445 level in HAECs transfected with sh-NC or sh-circ_0001445. (B-G) HAEC were treated with control, ox-LDL, ox-LDL+sh-NC, and ox-LDL+sh-circ_0001445. (B and C) ELISA was adopted for the secretions of TNF-α and IL-6 in treated HAECs. (D and E) Cell proliferative ability was assessed using CCK-8 assay and EdU assay in treated HAECs. (F) Migration ability was detected using Transwell assay in treated HAECs. (G) Protein levels of Collagen type II, MMP13, ADAMTS4 in treated HAECs were determined using western blot assay. **P <0.01, ***P <0.001.

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Yang, Z., Liang, X. & Yang, L. Circular RNA circ_0001445 alleviates the ox-LDL-induced endothelial injury in human primary aortic endothelial cells through regulating ABCG1 via acting as a sponge of miR-208b-5p. Gen Thorac Cardiovasc Surg 70, 779–792 (2022). https://doi.org/10.1007/s11748-022-01799-2

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