Shi Wenjie, Ren Changhong, Zhang Wei, Gao Chen, Yu Wantong, Ji Xunming, Chang Lisha
North China University of Science and Technology Affiliated Hospital, Tangshan 063000, China; Beijing Key Laboratory of Hypoxia Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
Beijing Key Laboratory of Hypoxia Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing 100053, China; Center of Stroke, Beijing Institute for Brain Disorder, Capital Medical University, Beijing 100053, China.
Neuroscience. 2023 Aug 21;526:35-47. doi: 10.1016/j.neuroscience.2023.06.009. Epub 2023 Jun 16.
Although hypoxic postconditioning (HPC) has a protective effect on ischemic stroke, its effect on angiogenesis after ischemic stroke is still unclear. This study was designed to investigate the effects of HPC on angiogenesis after ischemic stroke and to preliminarily study the mechanism involved. Oxygen-glucose deprivation (OGD)-intervened bEnd.3 (mouse brain-derived Endothelial cell. 3) was used to simulate cerebral ischemia. Cell counting kit-8 (CCK-8), Cell BrdU proliferation, wound healing, Transwell and tube formation assays were used to evaluate the effect of HPC on the cell viability, proliferation, migration (horizontal and vertical migration), morphogenesis and tube formation of bEnd.3. A middle cerebral artery occlusion (MCAO) model was made in C57 mice to simulate focal cerebral ischemia. Rod rotation test, corner test, modified neurological severity score (mNSS), and balance beam walking test were used to evaluate the effect of HPC on the neurological impairment of mice. Immunofluorescence staining was used to evaluate the effect of HPC on angiogenesis in mice. The angiogenesis-related proteins were evaluated and quantified using western blot. Results showed that HPC significantly promoted proliferation, migration and tube formation of bEnd.3. HPC significantly reversed the neurological deficit of MCAO mice. Moreover, HPC significantly promoted angiogenesis in the peri-infarct area, and angiogenesis was found to be positively correlated with the improvement of neurological impairment. The HPC mice showed higher PLCλ and ALK5 than did MCAO. We conclude that HPC improves the neurological deficit caused by focal cerebral ischemia by promoting angiogenesis. Furthermore, the effect of HPC on improving angiogenesis may be related to PLCλ and ALK5.
尽管缺氧后适应（HPC）对缺血性中风具有保护作用，但其对缺血性中风后血管生成的影响仍不清楚。本研究旨在探讨HPC对缺血性中风后血管生成的影响，并初步研究其相关机制。采用氧糖剥夺（OGD）干预的bEnd.3（小鼠脑源性内皮细胞3）来模拟脑缺血。使用细胞计数试剂盒-8（CCK-8）、细胞BrdU增殖、伤口愈合、Transwell和管形成实验来评估HPC对bEnd.3细胞活力、增殖、迁移（水平和垂直迁移）、形态发生和管形成的影响。在C57小鼠中制作大脑中动脉闭塞（MCAO）模型以模拟局灶性脑缺血。采用棒旋转试验、角落试验、改良神经功能缺损评分（mNSS）和平衡木行走试验来评估HPC对小鼠神经功能缺损的影响。采用免疫荧光染色来评估HPC对小鼠血管生成的影响。使用蛋白质免疫印迹法对血管生成相关蛋白进行评估和定量。结果显示，HPC显著促进了bEnd.3的增殖、迁移和管形成。HPC显著逆转了MCAO小鼠的神经功能缺损。此外，HPC显著促进了梗死灶周围区域的血管生成，并且发现血管生成与神经功能缺损的改善呈正相关。HPC小鼠的PLCλ和ALK5水平高于MCAO小鼠。我们得出结论，HPC通过促进血管生成改善局灶性脑缺血所致的神经功能缺损。此外，HPC改善血管生成的作用可能与PLCλ和ALK5有关。
