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  

Correlative Study on Calcium Influx and Expression of p53 Gene and Apoptosisin in Cerebral Cortical Cells of Neonatal Pigs Following Hypoxic Ischemic Encephalopathy.

作　　者：王晓蕾[1] 刘晓平[1] 刘凯[1] 张俊梅[1] 唐方[1] 余世禄[1] 

机构地区：[1]四川省成都市第三人民医院儿科,610031

出　　处：《中国儿童保健杂志》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)

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.

关 键 词：缺氧缺血性脑病 脑皮层细胞凋亡 细胞内游离钙 

分 类 号：R722.12[医药卫生—儿科]