PFT-α对脑缺血后神经干细胞移植中PUMA的调控研究  

Study on regulating of PFT-a to the PUMA in neural stem cells transplantation after cerebral ischemia

作　　者：刘洋[1] 雷旭辉[1] 殷实[1] 蒋传路[1] 

机构地区：[1]哈尔滨医科大学附属第二医院神经外科,150001

出　　处：《疑难病杂志》2014年第11期1156-1159,F0003,共5页Chinese Journal of Difficult and Complicated Cases

基　　金：黑龙江省科学基金资助项目(No.ZJY0604-02)

摘　　要：目的探索p53抑制剂(PET-α)对脑缺血后神经干细胞(NSCs)中p53下游促凋亡基因(PuMA)的表达影响。方法利用SD仔鼠分离NSCs,原代及传代培养,经氧糖剥夺(OGD)处理6 h后分为OGD+二甲基亚砜(DMSO)组和OGD+PFT-α组,采用免疫印迹技术比较2组p53蛋白及PUMA的表达变化。体内实验中采用Longa法建立经CT证实的大脑中动脉栓塞模型大鼠48只,随机分为PFT-α+NSCs移植组(n=24)和DMSO+NSCs移植组(n=24);采用免疫荧光技术检测不同移植组中PUMA的表达。结果 (1)体外实验中氧糖剥夺6 h后OGD+PFT-α组中p53蛋白入核水平下调,胞浆内表达上调,而OGD+DMSO组主要表达位于胞核;PUMA的表达水平明显低于OGD+DMSO组(P<0.05);(2)体内实验中,与DMSO+NSCs组相比,PIT-α+NSCs移植组中PUMA表达阳性的细胞明显减少[(38.9±4.3)%vs.(29.4±5.6)%,P<0.01]。结论 PFT-α与神经干细胞联合应用后,可明显抑制PUMA的表达,这可能是PFT-α促进移植后神经干细胞在脑缺血区存活的重要机制。Objective To explore the effect of p 53 inhibitor （ PFT-α） on neural stem cells after cerebral ischemia in p53 downstream apoptotic gene （PUMA） expressio.Methods Using SD rat to isolate NSCs, through the primary and pas-sage culture , by oxygen glucose deprivation （ OGD） treatment for 6 h, they were divided into OGD ＋dimethyl sulfoxide （ DM-SO） group and OGD＋PFT-αgroup, changes of expression of the p 53 protein and PUMA by immunoblotting technique were compared between the 2 group.In vivo tests using Longa method to establish the CT confirmed the brain artery embolism model in 48 rats, they were randomly divided into PFT-α＋NSCs transplantation group （ n =24） and DMSO＋NSCs trans-plantation group （ n =24） ;using immunofluorescence to detect the PUMA expression in the different groups of transplanta -tio.Results （1） In vitro oxygen glucose deprivation after 6 h, OGD＋PFT-αgroup’s p53 protein into the nucleus level de-creased , expression is up-regulated in the cytoplasm , while OGD＋DMSO group was mainly expressed in the cell nucleus;the expression level of PUMA was significantly lower than that in OGD ＋DMSO group （ P 〈0.05）; （2） In vivo experiment, compared with the DMSO ＋NSCs group , PFT-α＋NSCs PUMA transplantation group ’ s positive expression cells was signifi-cantly reduced [（3.9 ±.3）%v.（2.4 ±.6）%, P 〈0.01.Conclusion The PFT-αcombined with neural stem cells can inhibit the expression of PUMA , which may be an important mechanism of PFT-αto promote cell survival in the cerebral ischemic area after the transplantation of neural stem.

关 键 词：PFT-Α p53下游促凋亡基因 神经干细胞 脑缺血 凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]