大脑中动脉光化学栓塞模型的改良及免疫表型分析
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深圳市科技计划项目(GJHZ20200731095205016)


The Optimization of Photothrombotic Cerebral Artery Occlusion Model and the Poststroke Immune Phenotypic Analysis
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This work is supported by Shenzhen Science and Technology Program (GJHZ20200731095205016)

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    摘要:

    光照缺血模型是常用的脑卒中永久性缺血模型,其原理是:光敏感染料在光照射下被激活并产生活性氧自由基,生成的活性氧自由基会引起血管内皮损伤,导致血小板的黏附和聚集,并形成血栓。由于光照缺血模型产生的缺血半影带较小,同临床病理差异较大,限制了对于缺血半影带的研究,因此,该研究基于光化学法构建了一种改良的中动脉近端栓塞模型。具体地,向小鼠体内注射光敏感染料孟加拉红后,用激光照射中动脉近端 3 min,以诱导血管栓塞,并通过氯化三苯基四氮唑染色、免疫荧光以及流式细胞术对模型进行评估。氯化三苯基四氮唑染色和免疫荧光结果显示,与传统的光照缺血模型相比,该模型能在纹状区和皮质区产生较大且稳定的梗死区域(9%~15%),且能通过流式细胞术分离出梗死组织中的驻留小胶质细胞、浸润髓系细胞和淋巴细胞。上述实验结果表明,改良的中动脉近端光化学栓塞模型能够应用于缺血性脑卒中损伤后病理和免疫反应机制的研究。

    Abstract:

    Photothrombotic ischemia is a common experimental ischemic stroke model. In response to light stimulation, activated photosensitive dyes produce reactive oxygen species, which in turn induces damage to the vascular endothelial cells, causing platelet adhesion, aggregation and thrombosis. Since the conventional photothrombotic ischemia model produces only a tiny ischemic penumbra which can’t properly represent the clinical pathology, a modified proximal middle cerebral artery occlusion model was established in this study. The mouse proximal middle artery was irradiated by laser for 3 minutes to induce thrombosis following injection with the light-sensitive dye Rose Bengal and subsequently evaluated by 2,3,5-triphenyltetrazolium chloride staining, immunofluorescence, and flow cytometry. The results showed that this model produced a stable infarct area of 9% to 15% in the striatal and cortical regions, which is larger than the conventional photothrombotic ischemia. Resident microglia, infiltrating myeloid cells, and lymphocytes in the infarcted tissue were identified by flow cytometry. It is suggested that the modified proximal middle artery occlusion model can be applied to study the pathological and immune mechanisms after ischemic stroke injury.

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引文格式
徐帆,邓霞,许文,等.大脑中动脉光化学栓塞模型的改良及免疫表型分析 [J].集成技术,2023,12(2):10-19

Citing format
XU Fan, DENG Xia, XU Wen, et al. The Optimization of Photothrombotic Cerebral Artery Occlusion Model and the Poststroke Immune Phenotypic Analysis[J]. Journal of Integration Technology,2023,12(2):10-19

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  • 在线发布日期: 2023-03-23
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