CAJ | 학술논문

目的：探索p53抑制剂（PFT-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，PFT-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。
목적：탐색p53억제제（PFT-α）대뇌결혈후신경간세포（NSCs）중p53하유촉조망기인（PUMA）적표체영향。방법이용SD자서분리 NSCs，원대급전대배양，경양당박탈（ OGD）처리6 h후분위OGD＋이갑기아풍（ DMSO）조화OGD＋PFT-α조，채용면역인적기술비교2조p53단백급PUMA적표체변화。체내실험중채용Longa법건립경CT증실적대뇌중동맥전새모형대서48지，수궤분위PFT-α＋NSCs이식조（ n ＝24）화DMSO＋NSCs이식조（ n ＝24）；채용면역형광기술검측불동이식조중PUMA적표체。결과（1）체외실험중양당박탈6 h후OGD＋PFT-α조중p53단백입핵수평하조，포장내표체상조，이OGD＋DMSO조주요표체위우포핵；PUMA적표체수평명현저우OGD＋DMSO조（ P ＜0)．05）；（2）체내실험중，여DMSO＋NSCs조상비，PFT-α＋NSCs이식조중PUMA표체양성적세포명현감소[（38．9±4．3）％vs．（29．4±5．6）％， P ＜0．01]。결론 PFT-α여신경간세포연합응용후，가명현억제PUMA적표체，저가능시PFT-α촉진이식후신경간세포재뇌결혈구존활적중요궤제。
Objective To explore the effect of p 53 inhibitor ( PFT-α) on neural stem cells after cerebral ischemia in p53 downstream apoptotic gene (PUMA) expression.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 groups.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 -tion.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 [(38.9 ±4.3)%vs.(29.4 ±5.6)%, 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 .