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Knockdown of circRNA-Memo1 Reduces Hypoxia/Reoxygenation Injury in Human Brain Endothelial Cells Through miRNA-17-5p/SOS1 Axis

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Abstract

Circ-Memo1 has been proved to be upregulated in ischemia–reperfusion induced acute injury of kidney tissues. However, the potential role of circ-Memo1 in cerebral hypoxia/reoxygenation (H/R) injury is still unclear.

Blood samples were collected from 25 ischemic stroke patients and 25 healthy controls. To construct the H/R model, human brain microvascular endothelial cells (HBMVECs) were cultured under the hypoxic condition, followed by reoxygenation. Cell viability was analyzed by MTT assay. Flow cytometry was carried out to examine cell apoptosis. The level of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were measured by MDA and SOD assay kits, respectively. The levels of TNF-α, IL-1β, and IL-6 were determined by enzyme-linked immunosorbent assay (ELISA). Dual-luciferase reporter gene detection was employed to verify the binding relationships between circ-Memo1, miR-17-5p, and SOS1.

Circ-Memo1 and SOS1 expressions were increased, and miR-17-5p expression was reduced in ischemic stroke patients. Circ-Memo1 silencing promoted cell viability, inhibited the activation of ERK/NF-κB signaling pathway, reduced oxidative stress and inflammatory response, and inhibited cell apoptosis. Moreover, miR-17-5p functioned as the sponge of circ-Memo1, and SOS1 was identified as the target of miR-17-5p. The protective effect of circ-Memo1 knockdown on cell injury after H/R treatment was weakened by miR-17-5p inhibition.

Knockdown of circ-Memo1 alleviated H/R injury of HBMVEC cells by regulating the miR-17-5p/SOS1 axis, indicating that circ-Memo1 might be a potential treatment target for cerebral H/R injury.

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Availability of Data and Material

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Code Availability

Not applicable.

Abbreviations

CircRNAs:

Circular RNAs long non-coding RNAs

Circ-Memo1:

Circular mediator of ErbB2-driven cell motility 1

H/R:

Hypoxia-reoxygenation

IRI:

Ischemia-reperfusion injury

IRS1:

Insulin receptor substrate 1

HUVECs:

Human umbilical vein endothelial cells

HBMVECs:

Human brain microvascular endothelial cells

EMT:

Epithelial-mesenchymal transition

ERK:

Extracellular signal-regulating kinases

NF-κB:

Nuclear factor-κB

SOS1:

Son of Sevenless 1

ceRNA:

Competing endogenous RNA

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

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Authors and Affiliations

  1. Department of Neurology, Hunan Provincial People’s Hospitaland The First Affiliated Hospital of Hunan Normal UniversityHunan Province, No.569, Wangjiali Road, Changsha, 410016, People’s Republic of China

    Xiang Ren, Zhi-Wen Zhou & Jian-Wen Yang

  2. Department of Emergency, Hunan Provincial People’s Hospitaland The First Affiliated Hospital of Hunan Normal UniversityHunan Province, Changsha, 410016, People’s Republic of China

    Ying-Xia Jing

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Contributions

Conception and study design: XR.

Data acquisition: XR and YXJ.

Data analysis: ZWZ.

Funding acquisition: XR.

Manuscript drafting: XR and JWY.

Manuscript revising: XR.

All authors have read and approved the final version of this manuscript to be published.

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Correspondence to Xiang Ren.

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Ren, X., Jing, YX., Zhou, ZW. et al. Knockdown of circRNA-Memo1 Reduces Hypoxia/Reoxygenation Injury in Human Brain Endothelial Cells Through miRNA-17-5p/SOS1 Axis. Mol Neurobiol 59, 2085–2097 (2022). https://doi.org/10.1007/s12035-022-02743-4

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