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Knockdown of circHECTD1 inhibits oxygen-glucose deprivation and reperfusion induced endothelial-mesenchymal transition

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Abstract

Ischemic stroke (IS) has become a cerebrovascular disease which seriously threatens the elderly people. It has been reported that circRNAs participate in multiple diseases, including IS. However, the role of circHECTD1 in IS remains largely unknown. To mimic IS in vitro, human cerebral microvascular endothelial cells (HCMECs) were treated with oxygen glucose deprivation/reperfusion (OGD/R). Meanwhile, MCAO mouse model was established to detect the expression of circHECTD1 in IS. qRT-PCR and western blot were used to test gene and protein expressions, respectively. CCK-8 assay was used to investigate the cell viability. Moreover, cell migration and tube formation were assessed by transwell and tube formation assays. In addition, RIP and luciferase assay were performed to explore the association among circHECTD1, miR-335 and NOTCH2. CircHECTD1 was significantly upregulated in IS. OGD/R significantly induced EndoMT in HCMECs, while knockdown of circHECTD1 notably reversed this phenomenon. In addition, silencing of circHECTD1 remarkably reversed OGD/R-induced promotion of HCMEC tube formation and migration. Meanwhile, circHECTD1 upregulated the level of NOTCH2 through binding with miR-335. Furthermore, miR-335 inhibited the process of EndoMT in IS via targeting NOTCH2. In summary, circHECTD1 knockdown significantly alleviated EndoMT process in HCMECs via mediation of miR-335/NOTCH2 axis. Thus, circHECTD1 might act as a potential target against IS.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

Abbreviations

OGD/R:

Oxygen-glucose deprivation and reperfusion

IS:

Ischemic stroke

CircRNA:

Circular RNA

EndoMT:

Endothelial-to-mesenchymal transition

HECTD1:

HECT domain E3 ubiquitin ligase 1

MCAO:

Middle cerebral artery occlusion

HCMECs:

Human cerebral microvascular endothelial cells

TTC:

2,3,5-Triphenyltetrazolium chloride

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Authors and Affiliations

  1. Department of Neurology, The affiliated Changsha Central Hospital, Hengyang Medical School, University of South China, No.161 Shaoshan Road, Changsha, 410004, Hunan Province, China

    Guo-Hua He, Zhen Wang, Wei Xu, Kang-Ping Song & Hui Xiao

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  1. Guo-Hua He
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  2. Zhen Wang
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  3. Wei Xu
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  4. Kang-Ping Song
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  5. Hui Xiao
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Contributions

GHH conception, design of the work; HX, WX and KPS the acquisition, analysis and interpretation of data; ZW the creation of new software used in the work; GHH have drafted the work or substantively revised it; All authors read and approved the final manuscript.

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Correspondence to Guo-Hua He.

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This animal study was performed according to the guideline of National Institutes of Health and approved by the Ethics Committees of Changsha Central Hospital Institution.

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These authors declared no competing interests in this study.

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He, GH., Wang, Z., Xu, W. et al. Knockdown of circHECTD1 inhibits oxygen-glucose deprivation and reperfusion induced endothelial-mesenchymal transition. Metab Brain Dis 37, 427–437 (2022). https://doi.org/10.1007/s11011-021-00891-5

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