Yang Xinxuan, Zhao Jiahui, Tian Hao, Nie Ximing, Zheng Lina, Liu Xin, Wei Zheng Z, Ding Yuchuan, Liu Liping
Department of Neurology, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.
China National Clinical Research Center for Neurological Diseases, Beijing, China.
CNS Neurosci Ther. 2025 May;31(5):e70409. doi: 10.1111/cns.70409.
The no-reflow phenomenon in the ischemic brain following arterial recanalization leads to poor prognosis. Previous studies suggest that the clinically proven NA-1 drug, with disruption of PSD95 in the neuronal terminals alongside the cerebral microvasculature, may inhibit pericytic contraction of microvessels by reducing endothelin-1 secretion.
A 1.5-h tMCAO model using Balb/c mice was employed. In vivo two-photon imaging and immunofluorescence staining were employed to assess the constriction effect of pericytes on capillaries. The impact of NR2B9c/NA-1 administration (intravenously infused at a dose of 10 μmol/kg) on pericyte constriction was evaluated through immunofluorescent staining of brain sections. Additionally, The effect of NA-1 on cerebral perfusion was assessed using laser speckle blood flow monitoring, whole brain slice perfusion, and high-magnification capillary imaging. Enzyme-linked immunosorbent Assay (ELISA) was conducted to determine changes in quantitative ONOO and endothelin-1 (ET-1) content after tat-NA-1 administration. Lastly, microtubule-associated protein 2 (MAP2) staining and behavioral scores were used to evaluate the effects of NA-1 on infarct size and behavioral deficits.
In vivo two-photon imaging and immunofluorescence staining revealed that pericyte constriction following ischemia and recanalization resulted in a decreased diameter of capillaries, particularly at the soma and adjacent areas. Notably, capillary obstruction was localized near pericytes. After administration of NA-1, the immunofluorescence staining section showed that the diameter of capillaries at pericytes' soma and in the adjacent parts increased. The ELISA results indicated a reduction in ONOO and ET-1 levels. Additionally, MAP2 staining revealed a decrease in infarct size, while behavioral scores showed improvements in deficits. This effect of NA-1 was counteracted notably when added ET-1.
NA-1 inhibits pericyte constriction following ischemia-reperfusion by reducing ET-1 levels. It improves capillary no-reflow in mice, enhances cerebral perfusion, decreases infarct size, and mitigates behavioral deficits.
动脉再通后缺血性脑内的无复流现象会导致预后不良。先前的研究表明，临床验证的NA-1药物可破坏脑微血管旁神经元终末中的PSD95，可能通过减少内皮素-1分泌来抑制微血管周细胞的收缩。
采用Balb/c小鼠建立1.5小时大脑中动脉闭塞（tMCAO）模型。运用体内双光子成像和免疫荧光染色评估周细胞对毛细血管的收缩作用。通过脑切片免疫荧光染色评估NR2B9c/NA-1给药（以10 μmol/kg的剂量静脉输注）对周细胞收缩的影响。此外，使用激光散斑血流监测、全脑切片灌注和高倍毛细血管成像评估NA-1对脑灌注的影响。进行酶联免疫吸附测定（ELISA）以确定tat-NA-1给药后定量ONOO和内皮素-1（ET-1）含量的变化。最后，使用微管相关蛋白2（MAP2）染色和行为评分来评估NA-1对梗死面积和行为缺陷的影响。
体内双光子成像和免疫荧光染色显示，缺血再通后周细胞收缩导致毛细血管直径减小，尤其是在胞体和相邻区域。值得注意的是，毛细血管阻塞位于周细胞附近。给予NA-1后，免疫荧光染色切片显示周细胞胞体及其相邻部位的毛细血管直径增加。ELISA结果表明ONOO和ET-1水平降低。此外，MAP2染色显示梗死面积减小，而行为评分显示缺陷有所改善。当添加ET-1时，NA-1的这种作用明显被抵消。
NA-1通过降低ET-1水平抑制缺血再灌注后的周细胞收缩。它改善了小鼠的毛细血管无复流，增强了脑灌注，减小了梗死面积，并减轻了行为缺陷。
