ZEB2, interacting with MDM2, contributes to the dysfuntion of brain microvascular endothelial cells and brain injury after intracerebral hemorrhage

ABSTRACT

ZEB2 has been shown to be upregulated in the brain tissues of rats with intracerebral hemorrhage (ICH), but its role in ICH-caused brain injury remains unclear. Here, an ICH rat model was established via intracerebral injection of autologous blood, and the lentivirus-mediated ZEB2 short hairpin RNA (sh-ZEB2) or negative control (scramble) were administered 0.5 hours after ICH. Silencing ZEB2 alleviated ICH-induced neurologic deficits and the increase of BBB permeability, brain water content and ZEB2 expression. Next, OGD (oxygen glucose deprivation) plus hemin was used to treat primary brain microvascular endothelial cells (BMECs) to simulate the ICH condition in vitro. OGD plus hemin upregulated ZEB2 expression and apoptosis, but reduced cell viability, migration, TEER (transendothelial electric resistance) and the expression of vascular-endothelial (VE-) cadherin, occludin and claudin-5, which was reversed by inhibiting ZEB2. Mechanism researches showed that ZEB2 interacted with MDM2 to up-regulate MDM2 protein expression, and then increased E2F1 protein level by suppressing its ubiquitination, which in turn promoted the transcription of ZEB2 to induce its protein expression, so as to enhance the interaction between ZEB2 and MDM2, thereby contributing to OGD plus hemin-induced endothelial dysfunction. Additionally, the joint interference of ZEB2 and MDM2 in vivo had better mitigative effects on ICH-induced brain injury compared with silencing ZEB2 alone. In summary, ZEB2 interacted with MDM2 to promote BMEC dysfunction and brain damage after ICH.

KEYWORDS:

• Intracerebral hemorrhage
• ZEB2
• MDM2
• E2F1
• brain microvascular endothelial cells

Availability of data and materials

All data used during the current study are available from the corresponding author on reasonable request.

Credit authorship contribution statement

Qingbao Guo: Conceptualization, Investigation, Methodology, Writing - original draft. Manli Xie: Conceptualization, Investigation, Methodology, Writing - original draft. Miao Guo: Methodology, Software, Visualization. Feiping Yan: Resources, Validation, Data curation, Visualization. Lihong Li: Resources, Validation, Data curation, Software. Rui Liu: Conceptualization, Supervision, Writing - Review & Editing.

Disclosure statement

The authors declare that they have no competing interests.

Ethical approval

This research was approved by the Ethics Committee of Xing Yuan Hospital of Yulin (YLXY-2019-031). All animal experiments were in accordance with the guide for the care and use of laboratory animals established by United States National Institutes of Health (Bethesda, MD, USA).

Supplementary material

Supplemental data for this article can be accessed here