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Circ_0003423 Alleviates ox-LDL-Induced Human Brain Microvascular Endothelial Cell Injury via the miR-589-5p/TET2 Network

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Abstract

Brain microvascular endothelial cells (BMECs) injury is one of the main causes of cerebrovascular diseases. Circular RNA (circRNA) has been found to be involved in the regulation of cerebrovascular diseases progression. However, the role and mechanism of circ_0003423 in cerebrovascular diseases is still unclear. In our study, oxidized low density lipoprotein (ox-LDL)-induced HBMEC-IM cells were used to construct cerebrovascular cell injury model in vitro. Quantitative real-time PCR was used to determine the expression levels of circ_0003423, miR-589-5p and Ten-eleven translocation 2 (TET2). The interactions between miR-589-5p and circ_0003423 or TET2 were confirmed by dual-luciferase reporter assay, RIP assay and RNA pull-down assay. Cell viability, angiogenesis and apoptosis were measured using cell counting kit 8 assay, tube formation assay and flow cytometry. Cell oxidative stress was evaluated by detecting the levels of reactive oxygen species and lactate dehydrogenase. The protein levels were examined by western blot analysis. Our results showed that circ_0003423 was a downregulated circRNA in ox-LDL-induced HBMEC-IM cells. In the terms of mechanism, circ_0003423 was found to be a sponge of miR-589-5p. Function analysis showed that circ_0003423 overexpression could relieve ox-LDL-induced HBMEC-IM cell injury, and this effect could be reversed by miR-589-5p mimic. In addition, TET2 was confirmed to be a target of miR-589-5p, and its overexpression could alleviate ox-LDL-induced HBMEC-IM cell injury. Moreover, the rescue experiments also confirmed that TET2 silencing could abolish the inhibition effect of anti-miR-589-5p on ox-LDL-induced HBMEC-IM cell injury. In summary, our data showed that circ_0003423 alleviated ox-LDL-induced HBMEC-IM cells injury through regulating the miR-589-5p/TET2 axis.

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Funding

This work was supported by The Natural Science Foundation of Hainan Province (820MS143), Medical and Health Research Projects in Hainan Province (20A200364) and In-hospital Scientific Research and Cultivation Fund of the Second Affiliated Hospital of Hainan Medical College (The Second Affiliated Hospital of Haiyi Hospital (No. 50)).

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Author notes

  1. Hang Yu and Yuxiang Pan contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiovascular Surgery Intensive Care Unit, The Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan, China

    Hang Yu

  2. Department of Critical Care Medicine, The Second Affiliated Hospital of Hainan Medical University, No. 368 Yehai Avenue, Longhua District, Haikou, Hainan, China

    Yuxiang Pan, Huihui Xu & Jinguo Li

  3. Department of Neurology Three Areas, The Second Affiliated Hospital of Hainan Medical University, Haikou, Hainan, China

    Mingming Dai

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  1. Hang Yu
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  2. Yuxiang Pan
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  4. Huihui Xu
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  5. Jinguo Li
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Correspondence to Huihui Xu or Jinguo Li.

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Yu, H., Pan, Y., Dai, M. et al. Circ_0003423 Alleviates ox-LDL-Induced Human Brain Microvascular Endothelial Cell Injury via the miR-589-5p/TET2 Network. Neurochem Res 46, 2885–2896 (2021). https://doi.org/10.1007/s11064-021-03387-x

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  • DOI: https://doi.org/10.1007/s11064-021-03387-x

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