大鼠慢性局灶性脑低灌注模型建立及神经细胞凋亡的动态观测

发布时间：2018-08-31 08:59

【摘要】：目的: 本课题利用改良大鼠线栓法致MCAO建立慢性局灶性脑低灌注模型,研究慢性局灶性低灌注条件下神经细胞形态学的改变及血清hs-CRP的变化,为研究脑血管疾病及神经退行性疾病提供基础实验依据。 方法: 1、成年健康SD雄性大鼠60只,随机平均分为正常对照组、假手术组及造模后后1 d、3 d、7 d、14 d共6组,通过改良线栓法MCAO模型。应用神经行为学评分、TTC染色观察脑片及光镜、透射电镜观察细胞形态学的改变。 2、采用实验一所用大鼠。应用免疫组化及ELISA方法,观察rCCH大鼠不同时间点脑组织神经细胞凋亡的表达及血清中hs-CRP的含量。 结果: 1、各组大鼠术后两种神经行为学评分均≤1分,各组相互比较差异无统计学意义(P0.05);对照组与实验组大鼠脑切片TTC染色均未见局灶性脑梗灶。电镜可见神经细胞胞浆内细胞器崩解。 2、血清hs-CRP含量和神经细胞凋亡检测:术后1 d、3 d、7 d时,rCCH模型大鼠血清中hs-CRP含量与正常对照组比较,差异无显著性(P0.05),术后14 d时,rCCH模型大鼠血清中hs-CRP含量高于正常对照组,差异有显著性(P0.05);但hs-CRP数值在随日期递增,由此说明,在术后1-2 w时,随着实验时间的延长,rCCH模型大鼠血清中hs-CRP含量会继续增加。TUNEL法神经细胞凋亡染色显示,术后1 d内神经细胞凋亡细胞急剧增多,以星型胶质细胞为著,1 d组、3 d组及对照组间相比(P0.01);7 d到14 d间凋亡细胞依然呈增多趋势,但是由于时间的间隔,增加的数目相对减少。 结论: 1、通过改良大鼠线栓致MCAO模型,可成功建立较稳定的大鼠慢性局灶性低灌注模型。 2、慢性局灶性脑低灌注可导致供血区神经细胞过度凋亡,但未见片状炎性坏死。 3、rCCH大鼠MCA供血区神经细胞凋亡随术后时间的延长而逐渐加重。
[Abstract]:Objective: to establish a chronic focal cerebral hypoperfusion model by using modified rat thread occlusion method to study the morphological changes of nerve cells and the changes of serum hs-CRP in chronic focal cerebral hypoperfusion. To provide basic experimental basis for the study of cerebrovascular diseases and neurodegenerative diseases. Methods: 1. Sixty adult healthy male SD rats were randomly divided into normal control group, sham operation group and 6 groups at 1 day, 3 days and 7 days and 14 days after modeling. The MCAO model was established by modified suture method. The neurobehavioral score and TTC staining were used to observe the brain slices and light microscope, and the morphological changes were observed by transmission electron microscope. The expression of neuronal apoptosis and the content of hs-CRP in serum of rCCH rats at different time points were observed by immunohistochemical and ELISA methods. Results: 1. The scores of two kinds of neurological behavior were 鈮，

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