脑缺血后处理对大鼠大脑海马区Notch1、Cyclin D1和CDK4蛋白表达的影响

发布时间：2018-04-18 05:20

本文选题：缺血后处理 + Notch　；参考：《吉林大学学报(医学版)》2017年06期

【摘要】：目的:探讨基于Notch1信号通路观察缺血后处理(CIP)对全脑缺血再灌注(I/R)大鼠海马区神经元中Cyclin D1和CDK4蛋白表达的影响,阐明其机制。方法:128只健康的雄性SD大鼠随机分为假手术组、I/R组(改良的Pulsinelli四血管阻断法制作)、CIP组(在彻底再灌注之前进行反复3次的再通/阻断血管)和DAPT组(实施CIP之前3h按10mg·kg-1·d-1腹腔注射DAPT),每组32只。分别在缺血后6、24、48和72h采用HE染色观察各组大鼠海马区神经元表现并计算存活率,免疫组织化学染色观察Cyclin D1、CDK4和Notch1表达的细胞,Western blotting法观察各时间点大鼠海马区中Cyclin D1、CDK4和Notch1蛋白的表达水平。结果:HE染色,与假手术组比较,I/R组大鼠海马区神经元结构损伤,各时间点海马区神经元存活率明显降低(P0.05);与I/R组比较,CIP组相应时间点大鼠海马区神经元存活率明显升高(P0.05);与CIP组比较,DAPT组相应时间点大鼠海马区神经元存活率明显降低(P0.05)。免疫组织化学染色,与假手术组比较,I/R组大鼠海马区Notch1、Cyclin D1和CDK4阳性细胞数增加(P0.05);与I/R组比较,CIP组大鼠海马区Cyclin D1和CDK4阳性细胞数增加(P0.05),Notch1阳性细胞数明显增加(P0.05);与CIP组比较,DAPT组大鼠海马区Cyclin D1和CDK4阳性细胞数增加(P0.05),Notch1阳性细胞数明显减少(P0.05)。Western blotting法,与假手术组比较,I/R组大鼠海马区Notch1、Cyclin D1和CDK4蛋白表达水平升高(P0.05);与I/R组比较,CIP组大鼠海马区Cyclin D1和CDK4蛋白表达水平明显降低(P0.05),Notch1蛋白表达水平明显升高(P0.05);与CIP组比较,DAPT组的Cyclin D1和CDK4蛋白表达水平明显升高(P0.05),Notch1蛋白表达水平明显降低(P0.05)。结论:CIP对神经元的保护作用可能是通过提高Notch1蛋白活性和抑制Cyclin D1和CDK4蛋白实现的。
[Abstract]:Aim: to investigate the effects of Notch1 on the expression of Cyclin D1 and CDK4 in hippocampal neurons of rats with global cerebral ischemia-reperfusion (I / R), and to elucidate its mechanism.Methods Twenty eight healthy male Sprague-Dawley rats were randomly divided into two groups: sham operation group (I / R group) (modified Pulsinelli four-vessel occlusion method) and DAPT group (repeated recanalization / occlusion of blood vessels three times before reperfusion) and DAPT group (before CIP was performed).10mg kg-1 d-1 was injected intraperitoneally for 3 h with 32 rats in each group.The hippocampal neurons in each group were observed by HE staining and the survival rate was calculated at 6h 24 h and 72 h after ischemia, respectively.The expression levels of Cyclin D1tCDK4 and Notch1 protein in hippocampus of rats were observed by immunohistochemical staining. The expression of CDK4 and Notch1 in Cyclin D1 was detected by Western blotting method, and the expression of CDK4 and Notch1 in hippocampal area of rats at different time points were observed.Results compared with the sham-operated group, the hippocampal neuronal structure in the I / R group was damaged.Compared with I / R group, the survival rate of hippocampal neurons in CIP group was significantly higher than that in CIP group, and the survival rate of hippocampal neurons in DAPT group was significantly lower than that in CIP group at the corresponding time points.Immunohistochemical staining,Compared with the sham-operated group, the number of Notch1cyclin D1 and CDK4 positive cells in hippocampus increased in the I / R group, the number of Cyclin D1 and CDK4 positive cells in the hippocampal area of the I / R group increased significantly compared with that in the I / R group, and the number of P0.05 positive cells in the CIP group was significantly increased compared with that in the CIP group.The number of Cyclin D1 and CDK4 positive cells increased in equine area. The number of P0.05 Notch1 positive cells decreased significantly. Western blotting assay was used.Compared with the sham-operated group, the expression of Notch1Cyclin D1 and CDK4 protein in hippocampal area of rats in I- / R group was significantly higher than that in group I / R, and the expression level of Cyclin D1 and CDK4 protein in hippocampal area of rats in CIP group was significantly lower than that in group I / R, and the expression level of Notch1 protein in hippocampal area in group I / R was significantly higher than that in group I / R, and that in group CIP was significantly higher than that in group I / R.The expression level of Cyclin D1 and CDK4 protein in DAPT group was significantly higher than that in DAPT group.Conclusion the protective effect of Notch1 on neurons may be achieved by increasing the activity of Notch1 protein and inhibiting the expression of Cyclin D1 and CDK4 proteins.
【作者单位】：华北理工大学护理与康复学院;华北理工大学附属医院神经外科;河北省唐山市人民医院急诊科;
【基金】：河北省卫计委科研重点项目资助课题(ZD2010106)
【分类号】：R743

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