Xu Yiya, Chen Chao, Weng Jilin, Chen Ting, He Yingchao, Song Zhiwei, Wang Yinzhou
Shengli Clinical Medical College of Fujian Medical University, Fuzhou, Fujian, China.
Department of Neurology, Fuzhou University Affiliated Provincial Hospital, Fuzhou, Fujian, China.
Aging Dis. 2025 Jun 6. doi: 10.14336/AD.2025.0197.
Post-recanalization microcirculation dysfunction is common and significantly contributes to poor outcomes in ischemic stroke. Pericytes have been shown to mediate the "no-reflow" phenomenon by constricting capillaries in experimental stroke models, implicating their critical role in early microcirculation dysfunction. However, little is known about the long-term fate of pericytes and their contribution to sustained microcirculation dysfunction in prolonged period of time. We conducted repeated longitudinal observations of pericyte fate and function, as well as blood flow dynamics across multiple vascular segments, using two-photon imaging in PDGFRβ-tdTomato mice subjected to transient middle cerebral artery occlusion (tMCAO) over a 14-day period. Multivariate analysis was performed to identify imaging features independently associated with capillary perfusion on day 14. Types of pericyte death were assessed using immunohistochemistry and Western blot analysis. Fasudil and the RIPK1 inhibitor necrostatin-1 were administered to modulate pericyte dysfunction and survival during the acute and subacute phases of stroke. Outcomes were evaluated by total capillary perfusion, infarct volume, blood brain barrier (BBB) integrity, and neurological function over 14 days. Pericyte loss observed on day 7 post-stroke was independently associated with impaired microcirculation perfusion, as indicated by a reduction in total capillary volume. While fasudil treatment alone improved microcirculation perfusion on day 3, it did not alter pericyte fate or improve outcomes by day 14. Necroptosis was found to contribute to delayed pericyte loss in the ischemic penumbra. Combined therapy with fasudil and necrostatin-1 effectively prevented delayed pericytes loss and improved both microcirculation perfusion and neurological outcomes on day 14. Delayed pericyte loss contributes to irreversible microcirculation dysfunction in the subacute phase of stroke. Targeting pericyte dysfunction and necroptosis following recanalization represents a promising therapeutic strategy for enhance stroke recovery.
再通后微循环功能障碍很常见，并且显著导致缺血性卒中预后不良。在实验性卒中模型中，已证明周细胞通过收缩毛细血管介导“无复流”现象，这暗示了它们在早期微循环功能障碍中的关键作用。然而，关于周细胞的长期命运及其在较长时间段内对持续微循环功能障碍的作用知之甚少。我们使用双光子成像技术，在14天内对短暂性大脑中动脉闭塞（tMCAO）的PDGFRβ-tdTomato小鼠的周细胞命运和功能，以及多个血管段的血流动力学进行了重复的纵向观察。进行多变量分析以确定与第14天毛细血管灌注独立相关的成像特征。使用免疫组织化学和蛋白质印迹分析评估周细胞死亡类型。在卒中的急性期和亚急性期给予法舒地尔和RIPK1抑制剂坏死素-1，以调节周细胞功能障碍和存活。在14天内通过总毛细血管灌注、梗死体积、血脑屏障（BBB）完整性和神经功能评估结果。卒中后第7天观察到的周细胞损失与微循环灌注受损独立相关，表现为总毛细血管体积减少。虽然单独使用法舒地尔治疗在第3天改善了微循环灌注，但到第14天它并未改变周细胞命运或改善预后。发现坏死性凋亡导致缺血半暗带周细胞延迟损失。法舒地尔和坏死素-1联合治疗有效预防了周细胞延迟损失，并在第14天改善了微循环灌注和神经功能结局。延迟的周细胞损失导致卒中亚急性期不可逆的微循环功能障碍。再通后靶向周细胞功能障碍和坏死性凋亡是一种有前景的治疗策略，可促进卒中恢复。
