本文選題：缺血后處理 + Notch�。� 參考：《吉林大學學報(醫(yī)學版)》2017年06期

【摘要】：目的:探討基于Notch1信號通路觀察缺血后處理(CIP)對全腦缺血再灌注(I/R)大鼠海馬區(qū)神經(jīng)元中Cyclin D1和CDK4蛋白表達的影響,闡明其機制。方法:128只健康的雄性SD大鼠隨機分為假手術(shù)組、I/R組(改良的Pulsinelli四血管阻斷法制作)、CIP組(在徹底再灌注之前進行反復3次的再通/阻斷血管)和DAPT組(實施CIP之前3h按10mg·kg-1·d-1腹腔注射DAPT),每組32只。分別在缺血后6、24、48和72h采用HE染色觀察各組大鼠海馬區(qū)神經(jīng)元表現(xiàn)并計算存活率,免疫組織化學染色觀察Cyclin D1、CDK4和Notch1表達的細胞,Western blotting法觀察各時間點大鼠海馬區(qū)中Cyclin D1、CDK4和Notch1蛋白的表達水平。結(jié)果:HE染色,與假手術(shù)組比較,I/R組大鼠海馬區(qū)神經(jīng)元結(jié)構(gòu)損傷,各時間點海馬區(qū)神經(jīng)元存活率明顯降低(P0.05);與I/R組比較,CIP組相應時間點大鼠海馬區(qū)神經(jīng)元存活率明顯升高(P0.05);與CIP組比較,DAPT組相應時間點大鼠海馬區(qū)神經(jīng)元存活率明顯降低(P0.05)。免疫組織化學染色,與假手術(shù)組比較,I/R組大鼠海馬區(qū)Notch1、Cyclin D1和CDK4陽性細胞數(shù)增加(P0.05);與I/R組比較,CIP組大鼠海馬區(qū)Cyclin D1和CDK4陽性細胞數(shù)增加(P0.05),Notch1陽性細胞數(shù)明顯增加(P0.05);與CIP組比較,DAPT組大鼠海馬區(qū)Cyclin D1和CDK4陽性細胞數(shù)增加(P0.05),Notch1陽性細胞數(shù)明顯減少(P0.05)。Western blotting法,與假手術(shù)組比較,I/R組大鼠海馬區(qū)Notch1、Cyclin D1和CDK4蛋白表達水平升高(P0.05);與I/R組比較,CIP組大鼠海馬區(qū)Cyclin D1和CDK4蛋白表達水平明顯降低(P0.05),Notch1蛋白表達水平明顯升高(P0.05);與CIP組比較,DAPT組的Cyclin D1和CDK4蛋白表達水平明顯升高(P0.05),Notch1蛋白表達水平明顯降低(P0.05)。結(jié)論:CIP對神經(jīng)元的保護作用可能是通過提高Notch1蛋白活性和抑制Cyclin D1和CDK4蛋白實現(xiàn)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.
【作者單位】： 華北理工大學護理與康復學院;華北理工大學附屬醫(yī)院神經(jīng)外科;河北省唐山市人民醫(yī)院急診科;
【基金】：河北省衛(wèi)計委科研重點項目資助課題(ZD2010106)
【分類號】：R743

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