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Circ_0026218 ameliorates oxidized low-density lipoprotein-induced vascular endothelial cell dysfunction by regulating miR-188-3p/TLR4/NF-κB pathway

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Abstract

Background

Circular RNAs (circRNAs) have shown important regulatory roles in cardiovascular diseases, including atherosclerosis (AS). However, the role and mechanism of circ_0026218 in AS remain unclear.

Methods

The cell model of AS in vitro was established by stimulating human umbilical vein endothelial cells (HUVECs) with oxidized low-density lipoprotein (ox-LDL). In addition, circ_0026218, microRNA-188-3p (miR-188-3p), and toll-like receptor 4 (TLR4) expression was determined via real-time quantitative polymerase chain reaction (RT-qPCR) in serum samples from AS patients and healthy volunteers. Cell proliferation was assessed using Cell Counting Kit-8 (CCK-8) assay and 5-ethynyl-2’-deoxyuridine (EdU) assay. Cell apoptosis was measured using flow cytometry. The inflammatory response was assessed using enzyme-linked immunosorbent assay (ELISA). Oxidative stress level was assessed using corresponding kits. Nitric oxide (NO) level was examined using NO detection assay. The interaction between miR-188-3p and circ_0026218 or TLR4 was determined via dual-luciferase reporter, RNA immunoprecipitation (RIP), and RNA pull-down assays. Exosomes were observed using transmission electron microscopy (TEM). The size distribution of exosomes was analyzed using nanoparticle tracking analysis (NTA).

Results

Ox-LDL treatment caused HUVEC dysfunction by inhibiting cell proliferation and promoting apoptosis, inflammation, and oxidative stress. Circ_0026218 was upregulated in AS serum samples and ox-LDL-treated HUVECs. Knockdown of circ_0026218 attenuated ox-LDL-induced dysfunction in HUVECs. MiR-188-3p acted as a target of circ_0026218, and miR-188-3p downregulation reversed the suppression role of circ_0026218 knockdown on ox-LDL-induced HUVEC disorder. TLR4 was a target of miR-188-3p, and miR-188-3p overexpression alleviated ox-LDL-induced dysfunction in HUVECs by targeting TLR4. Circ_0026218 could deregulate the TLR4/NF-κB pathway by sponging the miR-188-3p. Importantly, circ_0026218 was overexpressed in exosomes from ox-LDL-treated HUVECs and could be delivered via exosomes.

Conclusion

Circ_0026218 knockdown attenuated ox-LDL-induced dysfunction in HUVECs via regulating miR-188-3p/TLR4/NF-κB pathway.

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Funding

Funding was provided by Natural Science Foundation of Xinjiang Uygur Autonomous Region (2018D01C219).

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Authors and Affiliations

  1. Departments of coronary heart disease, First Affiliated Hospital of Xinjiang Medical University, Urumqi City, China

    Jing Liu

  2. Departments of coronary heart disease, Xinjiang Medical University, Urumqi City, China

    Xiangyang Zhang

  3. Critical Care Medicine, First Affiliated Hospital of Xinjiang Medical University, Urumqi City, China

    Zhaoxia Yu

  4. Image Center, First Affiliated Hospital of Xinjiang Medical University, 137 Liushan South Road, Urumqi City, 830000, Xinjiang Province, China

    Tieliang Zhang

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Contributions

Xiangyang Zhang designed and performed the research; Zhaoxia Yu and Tieliang Zhang analyzed the data; Jing Liu wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Tieliang Zhang.

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ESM 1

Knockdown of circ_0026218 abolished ox-LDL-induced NO level repression in HUVECs. HUVECs were divided into four groups: Control (without ox-LD), ox-LDL (50 μg/mL), ox-LDL+si-NC, and ox-LDL+si-circ_0026218. NO level was assessed in HUVECs using NO detection assay (one-way ANOVA). **P < 0.01, ***P < 0.001. (PNG 13 kb)

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Liu, J., Zhang, X., Yu, Z. et al. Circ_0026218 ameliorates oxidized low-density lipoprotein-induced vascular endothelial cell dysfunction by regulating miR-188-3p/TLR4/NF-κB pathway. Cardiovasc Drugs Ther 38, 263–277 (2024). https://doi.org/10.1007/s10557-022-07416-x

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