新生大鼠痉挛型脑瘫动物模型构建及评价的实验研究

Experimental study on constructing and evaluating the neonatal rat model spastic cerebral palsy

目的:构建SCP动物模型,以期最大程度地模拟SCP的神经行为学及病理形态学改变,并探讨模型的评价方法。方法:清洁级5日龄W istar新生大鼠随机分为两组,实验组结扎双侧颈总动脉,而对照组仅切开颈部皮肤。检测方法分两部分:①两组各取20只新生大鼠,分别于术前、术后不同时期测量体重,术后25天进行神经行为学检测及数据统计分析;②两组各取16只新生大鼠,分别在术后不同时期对脑组织切片、HE染色及光镜观察。结果:①对照组存活率为100%,实验组存活率为85.71%。②术后实验组大鼠体重增长较对照组缓慢,差异有统计学意义(P<0.05)。③行为学检测结果显示,悬吊试验中、斜坡试验中、旷场试验中、拒俘反应试验中、肌张力测试中、不自主运动测试中,各组试验结果进行统计学分析后显示,均P<0.05,表明两组差异有统计学意义。④HE染色光镜观察,实验组脑组织病理改变为小胶质细胞浸润;星形胶质细胞肥大、增生;脑白质结构稀疏、紊乱;脑室周围白质软化灶形成,囊间少突胶质细胞减少;侧脑室扩大且形态多不规则等,对照组无异常。结论:①结扎5日龄新生大鼠双侧颈总动脉经神经行为学检测为成功构建痉挛型脑瘫动物模型;其脑损伤的部位主要在白质,该模型对脑瘫研究有重要意义。

Objective：Experimental study on constructing and evaluating the neonatal model of spastic cerebral palsy,to simulate the changes of neurology pathology on neonatal rats after SCP,and to explore the method of evaluating model further.Methods：The clean five-day-old Wistar neonatal rats were randomly divided into experimental and control group.Bilateral carotid ligation was used to perform on brain damage in 4 experimental group,while the neck skin was cut only in the control group.The experiment was carried out in two steps ： first,twenty rats in each group.Weight on postnatal first day（P6）,（P8）,（P12）and（P20） were measured,and their neurobehavior were tested on（P30）,to analyze the statistic data;second,sixteen neonatal rats in each group were chosed for the follow experiment： the pathological changes of brain tissue were observed under light microscope after HE staining at the time points of P6,P7,P15 and P25,respectively.Results：（1）The general situation after operation： the survival rate was 100%,in the control group,while survival rate was 85.71%.Depression,decreased limbs move-ments,dulled reaction and appetite loss were found in experiment rats;while the spirit of control rats recovered,limbs action remain active and sensitive and they fed gradually increased six hours later.（2）The body weight of two groups rats had no significant difference before surgery;The body weight of experimental rats showed slow growth,and there was significant difference at body weight aspect between experiment group and control groups after operation（P0.05）..（3）Pathological change： the pathological change of the model groups as follow①microglial cells infiltrated and aggregated in the white matter;②astrocytes were hypertrophic and proliferative;③white matter structure was sparse and disorderly;④periventricular leukomalacia lesions were formed and oligodendrocytes reduced between cysts;⑤lateral ventricle was enlarged and its shape was irregular;however,the pathology of the control rats wasn＇t observed abnormal in principle.（4）Neurobehavioral study changes： the average dropping time of experimental rats and in the control group,was statistical difference between the two groups（P0.05）;the average time of heading up in experimental rats and control group was significantly different（P0.05）;the average score of experimental and control group was statistically different（P0.05）.Conclusion：（1）Through the ligation on bilateral carotid artery with five-day-old neonatal rats,an spastic cerebral palsy model is successfully constructed.The main pathological change of spastic cerebral palsy model is in the white matter.The model is so practical to be used in cerebral palsy research.