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目的探讨经颈内动脉注射肺炎链球菌溶血素(PLY)建立感染性脑损伤动物模型的方法和可行性。方法采用颈内动脉注射PLY制作幼年大鼠感染性脑损伤动物模型。选取健康普通级1月龄SD大鼠160只,体质量100~120 g,雌雄不限,随机分为PLY组和对照组,其中PLY组(实验组)80只,颈内动脉注射PLY0.2 mL(7μg);对照组80只,颈内动脉注射等体积9 g.L-1盐水。根据不同观察时间点(注射后6 h、12 h、24 h及48 h),将PLY组SD大鼠分为4个亚组,每个亚组中分别有10只注射伊文思蓝(EB),测定不同时间点脑组织EB水平;注射EB组经右颈外静脉近心端注入20 g.L-1EB(2 mL.kg-1),推注时间1~2 min。各组处理后,待观察至相应时间点,将大鼠麻醉行心脏灌注后断头取脑,用于制作脑组织标本,以备观察。采用干湿质量法检测其脑组织含水量(BWC),甲酰胺法测定其脑组织EB水平,利用免疫组织化学法检测其脑组织神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)的表达。结果与对照组相比,HE染色光镜下观察,PLY组脑组织可见血管扩张、血管周围间隙增宽、炎细胞浸润、神经元空泡变性、星形胶质细胞体积增大肿胀、细胞核固缩等改变。各时间点PLY组BWC、EB水平、NSE及GFAP表达量均明显高于对照组,差异均有统计学意义(Pa<0.01)。结论经幼鼠颈内动脉注射PLY可以成功建立幼鼠感染性脑损伤模型。
Objective To investigate the feasibility and feasibility of establishing an animal model of infectious brain injury by injection of pneumolysin (PLY) through the internal carotid artery. Methods The animal model of infectious brain injury in young rats was made by injection of PLY into the internal carotid artery. A total of 160 healthy 1-month-old SD rats weighing 100-120 g were randomly divided into PLY group and control group, of which 80 were in PLY group (experimental group), PLY0.2 mL (7μg); control group of 80, the internal carotid artery was injected with an equal volume of 9 gL-1 saline. According to different observation time points (6 h, 12 h, 24 h and 48 h after injection), SD rats in PLY group were divided into 4 subgroups, 10 in each subgroup were injected Evans blue (EB) The levels of EB in brain tissue were measured at different time points. Injection of 20 gL-1EB (2 mL.kg-1) was given proximal to the right external jugular vein in EB group. The injection time was 1-2 min. After each group was treated, until the corresponding time point, the rats were anesthetized underwent cardiac perfusion after decapitation to take brain for brain tissue samples for observation. The brain water content (BWC) was measured by wet and dry method. The EB level in brain tissue was measured by formamide method. The expression of neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) expression. Results Compared with the control group, the brain tissue of PLY group showed vasodilatation, widened perivascular space, infiltration of inflammatory cells, degeneration of neurons and vacuolization, enlargement and swelling of astrocytes, Reduce the change. At each time point, the expression of BWC, EB, NSE and GFAP in PLY group were significantly higher than those in control group (P <0.01). Conclusion Injecting PLY into the internal carotid artery of young rats successfully established the model of infectious brain injury in young rats.