College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, PR China; Jinhua Institute of Zhejiang University, Jinhua 321299, China.
College of Pharmaceutical Sciences, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, PR China.
Exp Neurol. 2024 Sep;379:114870. doi: 10.1016/j.expneurol.2024.114870. Epub 2024 Jun 17.
The pathophysiological features of acute ischemic stroke (AIS) often involve dysfunction of the blood-brain barrier (BBB), characterized by the degradation of tight junction proteins (Tjs) leading to increased permeability. This dysfunction can exacerbate cerebral injury and contribute to severe complications. The permeability of the BBB fluctuates during different stages of AIS and is influenced by various factors. Developing effective therapies to restore BBB function remains a significant challenge in AIS treatment. High levels of vascular endothelial growth factor (VEGF) in the early stages of AIS have been shown to worsen BBB breakdown and stroke progression. Our study aimed to investigate the protective effects of the VEGF receptor inhibitor Axitinib on BBB dysfunction and cerebral ischemia/reperfusion-induced injury.
BEnd3 cell exposed to oxygen-glucose deprivation (OGD) model was constructed to estimate pharmacological activity of Axitinib (400 ng/ml) on anti-apoptosis and pathological barrier function recovery. In vivo, rats were subjected to a 1 h transient middle cerebral artery occlusion and 23 h reperfusion (tMCAO/R) to investigate the permeability of BBB and cerebral tissue damage. Axitinib was administered through the tail vein at the beginning of reperfusion. BBB integrity was assessed by Evans blue leakage and the expression levels of Tjs claudin-5 and occludin.
Our research revealed that co-incubation with Axitinib enhanced the cell viability of OGD-insulted bEnd3 cells, decreased LDH leakage rate, and suppressed the expression of apoptosis-related proteins cytochrome C and Bax. Axitinib also mitigated the damage to Tjs and facilitated the restoration of transepithelial electrical resistance in OGD-insulted bEnd.3 cells. In vivo, Axitinib administration reduced intracerebral Evans blue leakage and up-regulated the expression of Tjs in the penumbra brain tissue in tMCAO/R rats. Notably, 10 mg/kg Axitinib exerted a significant anti-ischemic effect by decreasing cerebral infarct volume and brain edema volume, improving neurological function, and reducing pro-inflammatory cytokines IL-6 and TNF-α in the brain.
Our study highlights Axitinib as a potent protectant of blood-brain barrier function, capable of promoting pathological blood-brain barrier recovery through VEGF inhibition and increased expression of tight junction proteins in AIS. This suggests that VEGF antagonism within the first 24 h post-stroke could be a novel therapeutic approach to enhance blood-brain barrier function and mitigate ischemia-reperfusion injury.
急性缺血性脑卒中（AIS）的病理生理特征通常涉及血脑屏障（BBB）的功能障碍，其特征是紧密连接蛋白（Tjs）的降解导致通透性增加。这种功能障碍会加重脑损伤，并导致严重的并发症。BBB 的通透性在 AIS 的不同阶段波动，并受到多种因素的影响。开发有效的治疗方法来恢复 BBB 的功能仍然是 AIS 治疗中的一个重大挑战。AIS 早期高水平的血管内皮生长因子（VEGF）已被证明会加重 BBB 破裂和中风进展。我们的研究旨在探讨 VEGF 受体抑制剂 Axitinib 对 BBB 功能障碍和脑缺血/再灌注诱导损伤的保护作用。
构建 BEnd3 细胞氧葡萄糖剥夺（OGD）模型，以评估 Axitinib（400ng/ml）对抗细胞凋亡和病理性屏障功能恢复的药理活性。在体内，大鼠接受 1 小时短暂性大脑中动脉闭塞和 23 小时再灌注（tMCAO/R），以研究 BBB 的通透性和脑组织损伤。Axitinib 在再灌注开始时通过尾静脉给药。通过 Evans 蓝渗漏和紧密连接蛋白 Claudin-5 和 Occludin 的表达水平评估 BBB 完整性。
我们的研究表明，与 Axitinib 共孵育可增强 OGD 损伤的 bEnd3 细胞的细胞活力，降低 LDH 漏出率，并抑制凋亡相关蛋白细胞色素 C 和 Bax 的表达。Axitinib 还减轻了 Tjs 的损伤，并促进了 OGD 损伤的 bEnd.3 细胞中上皮间电阻的恢复。在体内，Axitinib 给药可减少 tMCAO/R 大鼠脑内 Evans 蓝渗漏，并上调缺血半影脑组织中 Tjs 的表达。值得注意的是，10mg/kg 的 Axitinib 通过减少脑梗死体积和脑水肿体积、改善神经功能以及降低大脑中的促炎细胞因子 IL-6 和 TNF-α，发挥了显著的抗缺血作用。
我们的研究强调了 Axitinib 作为血脑屏障功能的有效保护剂，通过抑制 VEGF 和增加紧密连接蛋白的表达，促进病理血脑屏障的恢复。这表明，在中风后 24 小时内使用 VEGF 拮抗剂可能是一种增强血脑屏障功能和减轻缺血再灌注损伤的新治疗方法。
