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新生猪HIE脑皮层细胞凋亡、钙内流及抑癌基因p53蛋白的相关性研究 被引量:1

Correlative Study on Calcium Influx and Expression of p53 Gene and Apoptosisin in Cerebral Cortical Cells of Neonatal Pigs Following Hypoxic Ischemic Encephalopathy.
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摘要 【目的】 探讨新生动物缺氧缺血性脑病( HIE) 脑皮层细胞凋亡的发生及其相关的诱导因素。【方法】 用琼脂电泳观察“DNA 梯带”的出现,用流式细胞术(FCM) 对脑皮层细胞凋亡及细胞内游离钙(IFCa) 、抑癌基因p53表达水平进行动态研究。【结果】 缺氧缺血后4hr 新生猪同侧额顶叶脑皮层细胞未见明显凋亡,72hr 后实验组凋亡率较对照组及尼莫通治疗组明显出现( n = 15 ,12 ,13 ;-x ±s= 27 .50 ±16 .2 ,3 .23 ±2 .93 ,9 .17 ±5 .76 ,F= 18 .756 4 ,P< 0 .000 1) ;HI 后4hr IFCa 及p53 基因阳性表达率明显增高( n = 12 ,12 ,12 ;-x ±s = 348 .96 ±39 .59 ,160 .98 ±37 .05 ,260 .18 ±66 .81 ,F= 41 .168 1 ,P< 0 .000 1 ;-x ±s = 6 .07 ±5 .55 ,1 .18 ±0 .53 ,3 .13 ±2 .18 ,F= 6 .076 7 ,P= 0 .005 7) ,且两者间呈显著正相关( r = 0 .497 7 ,P= 0 .002) ;HI 后72hr 凋亡率与4hr IFCa 水平呈显著正相关( r = 0 ? Objective To determine whether there is apoptosis in cerebral cortical of the neonates following hypoxic ischemic encephalopathy(HIE)and the correlative inducement. Method The' DNA Ladder' was observed by gel electrophoresis and the dynamic changes of cerebral apoptosis ratio(CAR)and the intracellular free calcium (IFCa)level and the expression of p53 gene were investigated via flow cytometry(FCM). Results We found that there was no obvious apoptosis in ipsilateral frontotemporal corticocerebrum 4hr later post Hl.The CAR 72hr later post Hl in experimental group was significantly higher than those in control group and in Nimotop group( n =15,12,13; ±s =27.50±16.20,3.23±2.93,9.17±5.76; F =18.756 4, P <0.000 1);IFCa level and the positive expression of p53 protien 4hr later post HI in experimental group increased obviously( n =12,12,12; ±s =348.96±39.59,160.98±37.05,260.81±66.81, F =41.168 1, P <0.000 1; ±s =6.07±5.55, 1.18±0.53, 3.13±2.18, F =6.0767, P =0.0057).An obvious correlation existed between IFCa level and expression of p53 gene 4hr later post Hl( r =0.497 7, P =0.002).A positive correlation between CAR 72hr later and IFCa level 4hr later post HI ( r =0.661 5, P <0.001)was found.The'DNA Ladder'was correponded to appearance of the apoptosis peak observed by FCM. Conclusion The apoptosis of cerbral cortical cells plays an important role in the delayed cerebral damage of the neonates following Hl,and that this apoptosis can be induced mainly by means of activation of calcium channel and proper usage of calcium channel blockers Nimotop can partially inhibit this apoptosis.Simultaneously,the damage of cellular DNA following HIE is also an important inducement of the apoptosis.
出处 《中国儿童保健杂志》 CAS 2000年第1期47-50,共4页 Chinese Journal of Child Health Care
基金 四川省卫生厅及成都市卫生局科研基金!资助 国家九五攻点课题!( 编号969040604)
关键词 缺氧缺血性脑病 脑皮层细胞凋亡 细胞内游离钙 Hypoxic ischemic encephalopathy(HIE) Cerebral cortical cell Apoptosis Intracellular free calcium Expression of p53 gene
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