双光子显微镜飞秒激光照射建立小鼠大脑皮质微梗死模型  

A mouse cerebral cortical microinfarct model induced by ultrashort laser irradiation wih two-photon microscopy

作　　者：史涛涛[1] 罗世坚[1] 唐超刚[1] 陈晓锋[1] 冯玉琨[1] 黄如训[2] 裴中[2] 李振东[1] 

机构地区：[1]中山大学附属第五医院神经科,珠海519000 [2]中山大学附属第一医院神经科,广州510080

出　　处：《国际脑血管病杂志》2017年第5期425-430,共6页International Journal of Cerebrovascular Diseases

摘　　要：目的验证双光子显微镜建立小鼠大脑皮质微梗死模型的可靠性并初步探讨其病理改变。方法17只雄性C57BL／6J小鼠随机分为微梗死组（n=11）和假手术组（n=6）。使用高速钻头磨薄直径约3mm的颅骨窗至解剖显微镜下清晰可见颅骨下小血管，再用双光子显微镜发射飞秒激光经此颅骨窗照射诱导选定的大脑皮质单个穿支动脉永久性闭塞。在模型制作后7d时采用HE染色法检测梗死体积，采用免疫组织化学法评价神经元死亡、胶质细胞活化和3-硝基酪氨酸（3-nitrotyroshae，3-NT）沉积。结果微梗死组有8只（72．7％）小鼠大脑皮质目标血管被闭塞并形成微梗死病灶，平均体积为（317．23±20．29）μm^3；梗死核心区可见大量神经元脱失和小胶质细胞浸润，梗死灶环绕着大量活化的星形胶质细胞，梗死周围区可见大量3-NT沉积。假手术组未见梗死灶，3-NT沉积显著低于微梗死组（8．00±1．479对98．38±9．10；t=23．962，P〈0．001）。结论双光子显微镜激光照射闭塞小鼠大脑皮质单个穿支动脉建立微梗死模型的方法可靠，脑组织病理学改变符合脑微梗死的特征。Objeelive To verify the reliability of the mouse model of cerebral cortical microinfarct induced by two-photon microscopy and to explore its pathological changes. Methods Seventeen male C57BL/6J mice were randomly divided into a microinfarct group （n = 11） or a sham operation group （n =6）. A thinned cranial window of 3 mm diameter was performed over the cerebral cortex with a high-speed micro-driU until the small blood vessels were clearly observed under a dissecting microscope. Then, a permanent single cortical penetrating arteriole occlusion was induced with a gradually enhanced ultrashort laser irradiation through the thinned cranial window with two-photon microscopy. At 7 days after modeling, the cerebral microinfarct volume was measured with HE staining, and the neuron loss, activation of glial cells and deposition of 3-nitrotyrosine were assessed using immunohistochemistry. Resulls The target vessels of cerebral cortex in 8 （72.7%） mice were occluded and the microinfarcts formed in the microinfarct group, and the average microinfarct volume was 317.23 ± 20.29 μm3. There were remarkable neuron loss and microglia infiltration in the infarcted core, a large number of reactive astrocytes surrounding the infarcted lesion, and massive deposition of 3-nitrotyrosine in the peri-infarct area. No infarcts were observed in the sham operation group. The deposition of 3-nitrotyrosine in the sham operation group was significantly less than that in the microinfarct group （8. 00 ± 1.48 vs. 98.38 ± 9. 10; t =23. 962, P 〈 0. 001）. Conclusions The mouse model of cerebral cortical microinfarct induced by two-photon microscopy is reliable, and its histopathologic changes are consistent with the pathologic features of cerebral microinfarct.

关 键 词：脑梗死 大脑皮质 显微术 共焦 神经元 细胞死亡 炎症 酪氨酸 疾病模型 动物 小鼠 

分 类 号：R-332[医药卫生] R743.3