结扎颈总动脉复合低氧致C 57 BL/6小鼠缺血缺氧性脑病模型的建立及溶栓胶囊的作用

Establishment of C 57 BL/6 Mice of Hypoxic-ischemic Encephalopathy(HIE) Model by Arteria Carotis Communis Ligation Combined with Hypoxia Inhalation and Effects of Rongshuan Capsules

目的:通过结扎颈总动脉复合低氧建立C 57 BL/6小鼠缺血缺氧性脑病(HIE)的模型,并观察溶栓胶囊的作用。方法:C 57 BL/6小鼠随机分为假手术组、模型组、溶栓胶囊大、中、小剂量组(400、200、100 mg生药/kg)。采用左侧颈总动脉结扎并通入氧气和氮气(8∶92)15 min的方法,建立C 57 BL/6小鼠缺血缺氧性脑病模型。在缺血缺氧损伤24 h后开始灌胃给药,1次/d,连续给药4周。4周后观察模型小鼠肌力、学习记忆能力的改变以及脑组织病理形态学变化以及NSE、MAP-2和Caspase-3的表达。结果 :手术后4周,缺血缺氧性脑病C 57 BL/6小鼠肌力和学习记忆能力显著下降,NSE和MAP-2低表达,Caspase-3高表达,病理学检查可见神经元细胞排列紊乱,大量变性、坏死及凋亡细胞,另见大片软化灶,血管扩张充血,多量炎性反应细胞浸润。溶栓胶囊对上述指标有明显改善作用。结论:通过结扎颈总动脉合并低氧成功建立了C 57 BL/6小鼠缺血缺氧性脑病模型,溶栓胶囊可改善模型小鼠肌力、学习记忆下降等症状,减轻脑组织病理性损伤,其机制可能与其神经保护作用有关。

Objective： To establish hypoxic-ischemic encephalopathy（ HIE） model on C 57 BL/6 mice by carotid artery ligation and hypoxia inhalation,and to observe the effect of Rongshuan Capsules. Methods： C 57 BL/6 mice were randomly divided into5 groups： sham-operation group,model group,large dosage of Rongshuan Capsules,middle dosage of Rongshuan Capsules,and small dosage of Rongshuan Capsules（ 400 mg/kg,200 mg/kg and 100 mg/kg）. HIE model mice were established by left carotid artery ligation on C 57 BL/6 mice and hypoxia inhalation（ O2： N2= 8 ∶ 92） for 15 minutes. Intragastric administration was used on HIE model mice 24 hours after ischemia anoxic damage,1 time/day and consecutively treated for 4 weeks. After 4 weeks of treatment,muscle strength,ability of learning and memory,NSE,MAP-2 and Caspase-3 expression and the brain tissue pathomorphological changes were tested. Results： Four weeks after model establishment,muscle strength and learning and memory ability of HIE model mice were decreased significantly. MAP-2 and NSE expression was down-regulated,and Caspase-3 expression was upregulated. Pathological examination showed that neurons were arranged in disorders,large amount of degenerated,necrotic and apoptotic cells. Large pieces of softening lesion,expansion and congestion of vessels and inflammatory cell infiltration were also observed. Rongshuan Capsules improved these indexes significantly in drug administration groups. Conclusion： Caroted artery ligation with hypoxia successfully established HIE models on C 57 BL/6 mice. Rongshuan Capsules facilitated model mice of muscle strength and learning and memory ability as well as MAP-2 and Caspase-3 expression,release the pathological injury of brain tissues,whose mechanism may be related to its neural protection.