Department of Biomedical Sciences, Ajou University Graduate School of Medicine, Suwon, South Korea.
Department of Neurology, Ajou University School of Medicine, Ajou University Medical Center, Suwon, South Korea.
Neurobiol Dis. 2019 Dec;132:104538. doi: 10.1016/j.nbd.2019.104538. Epub 2019 Jul 22.
It is challenging to revitalize ischemic penumbra after an acute stroke with intracranial perfusion insufficiency. To evaluate whether cranial burr hole and erythropoietin (EPO) generate effective revascularization, we investigated the efficacy of the augmentation method for reverse arteriogenesis from the healthy extracranial milieu. An intracranial perfusion insufficiency was created through bilateral internal carotid artery ligation (bICAL) in Sprague-Dawley rats. We administered recombinant human EPO (5000 U/kg) or saline intraperitoneally for 3 days after bICAL. Mechanical barrier disruption (MBD) was performed through a cranial burr hole with small dural cracks in the right hemisphere. The ipsilateral hemisphere with MBD grossly showed vascular networks between the extra- and intra-cranial spaces 2 weeks after the MBD procedure. It also showed significantly increased vessels in the intracranial vasculature adjacent to the MBD region (p = 0.0006). The levels of pro-angiogenic and inflammatory factors with prominent markers of vessel permeability were also significantly increased (MBD-only vs. control; Tnf-α, p = 0.0007; Vegf, p = 0.0206). In the EPO-administered group, such elevations in inflammation were significantly mitigated (combined vs. MBD-only; Tnf-α, p = 0.0008). The ipsilateral hemisphere with MBD-EPO (vs. MBD-only) showed significantly increased vessels (RECA-1, p = 0.0182) and their maturation (RECA-1/α-SMA, p = 0.0046), with upregulation of tumor growth factor-β1 (Tgf-β1, p = 0.037) and matrix metalloproteinase-2 (Mmp-2, p = 0.0488). These findings were completely blocked by minocycline (MIC) administration during in vivo (Tgf-β1, p = 0.0009; Mmp-2, p < 0.0001) and in vitro experiments (tube formation, p < 0.0001). Our data suggest that the MBD procedure (for angiogenic routes) and EPO administration (for an arteriogenic booster) are complimentary and can facilitate successfully "reverse arteriogenesis" in subjects with intracranial perfusion insufficiency.
在颅内灌注不足的急性脑卒中后，使缺血半影区重新恢复活力具有挑战性。为了评估颅骨钻孔和促红细胞生成素（EPO）是否能有效实现再血管化，我们研究了从健康的颅外环境增强逆行动脉生成的扩增方法的疗效。通过双侧颈内动脉结扎（bICAL）在 Sprague-Dawley 大鼠中创建颅内灌注不足。在 bICAL 后，我们通过右半球颅骨钻孔和小硬脑膜裂缝进行机械屏障破坏（MBD）。MBD 后 2 周，MBD 手术同侧半球在颅内血管附近的 MBD 区域明显增加了血管（p=0.0006）。MBD 区域还显著增加了血管通透性的突出标志物的促血管生成和炎症因子的水平（MBD 仅与对照相比；Tnf-α，p=0.0007；Vegf，p=0.0206）。在 EPO 给药组中，炎症的这种升高明显减轻（联合与 MBD 仅相比；Tnf-α，p=0.0008）。与 MBD 仅相比，MBD-EPO 的同侧半球（MBD 仅）显示出明显增加的血管（RECA-1，p=0.0182）和它们的成熟（RECA-1/α-SMA，p=0.0046），肿瘤生长因子-β1（Tgf-β1，p=0.037）和基质金属蛋白酶-2（Mmp-2，p=0.0488）上调。这些发现通过米诺环素（MIC）在体内（Tgf-β1，p=0.0009；Mmp-2，p<0.0001）和体外实验（管形成，p<0.0001）期间给药完全阻断。我们的数据表明，MBD 程序（用于血管生成途径）和 EPO 给药（用于促动脉生成增强剂）是互补的，可以成功促进颅内灌注不足患者的“逆行动脉生成”。
