Baotou Vocational and Technical College, Baotou City, Inner Mongolia, China.
Bioengineered. 2022 Mar;13(3):7760-7770. doi: 10.1080/21655979.2022.2045833.
Ischemic stroke is a disease in which brain tissue is damaged by a sudden rupture or blockage of a blood vessel in the brain that prevents blood from flowing to the brain. Extensive literature has demonstrated the neuroprotective effect of donepezil on brain injury, and this paper attempts to further reveal the effect of donepezil on brain microvascular endothelial cells dysfunction. Human brain microvascular endothelial cells (HBMECs) were treated with oxygen-glucose deprivation/reoxygenation (OGD/R) to induced brain microvascular endothelial cell dysfunction. The OGD/R-induced cell were added with different doses of donepezil with or without Sirtuin-1 (SIRT1) inhibitor EX527. Cell viability of HBMECs was examined by cell counting kit (CCK)-8 assay. OGD/R-treated cell migration was assessed by wound healing assay while angiogenesis in HBMECs was examined by tube formation assay and Western blot analysis. Endothelial cell dysfunction was assessed employing fluorescein isothiocyanate-dextran assay and Western blotting. SIRTI/FOXO3a/NF-kB signaling pathway-related protein expressions were detected using Western blotting. After pretreatment with SIRT1 inhibitor EX527, the above experiments were done again. Donepezil increased cell viability of OGD/R-induced HBMECs, promoted cell migration and angiogenesis, decreased cell permeability, and upregulated the expressions of tight junction proteins. In addition, donepezil regulated the expressions of SIRT1/FOXO3a/NF-κB signaling pathways. However, pretreatment with the SIRT1 inhibitor EX527 reversed the protective effect of donepezil on OGD/R-induced HBMECs. In summary, Donepezil ameliorates OGD/R-induced brain microvascular endothelial cell dysfunction via the SIRT1/FOXO3a/NF-κB pathways.
缺血性脑卒中是一种疾病,其中脑组织因大脑血管突然破裂或阻塞而受损,导致血液无法流向大脑。大量文献已经证明了多奈哌齐对脑损伤的神经保护作用,本文试图进一步揭示多奈哌齐对脑微血管内皮细胞功能障碍的影响。用氧葡萄糖剥夺/复氧(OGD/R)处理人脑微血管内皮细胞(HBMEC)以诱导脑微血管内皮细胞功能障碍。用不同剂量的多奈哌齐处理 OGD/R 诱导的细胞,或用 Sirtuin-1(SIRT1)抑制剂 EX527 处理。通过细胞计数试剂盒(CCK)-8 测定 HBMEC 细胞活力。通过划痕愈合试验评估 OGD/R 处理细胞的迁移,通过管形成试验和 Western blot 分析评估 HBMEC 中的血管生成。通过荧光素异硫氰酸酯-葡聚糖测定评估内皮细胞功能障碍,通过 Western blot 分析评估 SIRTI/FOXO3a/NF-κB 信号通路相关蛋白的表达。用 SIRT1 抑制剂 EX527 预处理后,再次进行上述实验。多奈哌齐增加 OGD/R 诱导的 HBMEC 细胞活力,促进细胞迁移和血管生成,降低细胞通透性,并上调紧密连接蛋白的表达。此外,多奈哌齐调节 SIRT1/FOXO3a/NF-κB 信号通路的表达。然而,用 SIRT1 抑制剂 EX527 预处理会逆转多奈哌齐对 OGD/R 诱导的 HBMEC 的保护作用。总之,多奈哌齐通过 SIRT1/FOXO3a/NF-κB 通路改善 OGD/R 诱导的脑微血管内皮细胞功能障碍。