Cell Cycle ( IF 4.3 ) Pub Date : 2021-08-02 , DOI: 10.1080/15384101.2021.1959702 Qingbao Guo 1 , Manli Xie 2 , Miao Guo 3 , Feiping Yan 4 , Lihong Li 1 , Rui Liu 5
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.
中文翻译:
ZEB2 与 MDM2 相互作用导致脑微血管内皮细胞功能障碍和脑出血后脑损伤
摘要
ZEB2 已被证明在脑出血 (ICH) 大鼠的脑组织中上调,但其在 ICH 引起的脑损伤中的作用仍不清楚。在这里,通过脑内注射自体血建立了 ICH 大鼠模型,并在 ICH 后 0.5 小时给予慢病毒介导的 ZEB2 短发夹 RNA (sh-ZEB2) 或阴性对照 (scramble)。沉默 ZEB2 可减轻 ICH 引起的神经功能缺损和 BBB 通透性、脑含水量和 ZEB2 表达的增加。接下来,OGD(氧葡萄糖剥夺)加血红素用于治疗原代脑微血管内皮细胞(BMECs)以模拟体外ICH情况. OGD 加血红素上调 ZEB2 表达和凋亡,但降低细胞活力、迁移、TEER(跨内皮电阻)和血管内皮 (VE-) 钙粘蛋白、occludin 和 claudin-5 的表达,通过抑制 ZEB2 可逆转。机制研究表明,ZEB2与MDM2相互作用上调MDM2蛋白表达,进而通过抑制其泛素化提高E2F1蛋白水平,进而促进ZEB2转录,诱导其蛋白表达,从而增强ZEB2与MDM2,从而导致 OGD 加血红素诱导的内皮功能障碍。此外,ZEB2 和 MDM2在体内的联合干扰与单独沉默 ZEB2 相比,对 ICH 引起的脑损伤有更好的缓解作用。总之,ZEB2 与 MDM2 相互作用促进 ICH 后的 BMEC 功能障碍和脑损伤。