抑制p53通路促进移植神经干细胞在脑缺血区的存活  

Inhibition of p53 pathway enhances the survival of grafted neural stem cells in ischemia regions of brain

作　　者：雷旭辉[1] 张平[1] 蒋传路[1] 杜文众[1] 叶伟[1] 

机构地区：[1]哈尔滨医科大学附属第二医院神经外科,150086

出　　处：《中国微侵袭神经外科杂志》2014年第3期129-131,共3页Chinese Journal of Minimally Invasive Neurosurgery

基　　金：黑龙江省卫生厅资助项目(编号:2013047);黑龙江省科学基金资助项目(编号:ZJY0604-02);哈尔滨市科技创新人才研究专项资金项目(编号:2010RFXXS021)

摘　　要：目的探讨一种提高移植神经干细胞(neural stem cells,NSC)在脑缺血区存活的方法。方法原代培养胚胎NSC并鉴定。采用改良Longa线栓法成功制作60只大鼠大脑中动脉阻塞模型,随机分为3组:对照组(n=12)、NSC移植组(n=24)和NSC+P53抑制剂(pifithrin-α,PFT-α)移植组(n=24)。以上3组在脑梗死模型建立24 h后,分别于脑内特定部位注射等量DMEM/F12培养液、NSC悬液和NSC+PFT-α悬液。细胞移植12 h、24 h和7 d后行免疫荧光染色,检测P53蛋白表达和移植NSC的存活情况。结果移植后12 h,NSC移植组P53蛋白主要表达在移植细胞的细胞核,而NSC+PFT-α移植组P53蛋白的表达主要位于移植细胞的细胞质。移植后7 d,NSC+PFT-α移植组NSC的存活数为(18.20±2.69)/高倍镜视野,NSC移植组为(13.06±3.20)/高倍镜视野,两组间比较差异具有统计学意义(P<0.05)。结论 NSC移植联合PFT-α对提高移植细胞的存活率有重要意义。Objective To investigate a method for increasing the survival rate of neural stem cells （NSC） after transplantation in ischemia regions of brain. Methods Primary NSC obtained from brain tissue of fetal SD rats were cultured and identified. The middle cerebral artery occlusion model was successfully established in 60 SD rats by modified Longa suture method, which were randomly divided into control group （n = 12）, NSC transplantation group （n = 24） and NSC＋P53 inhibitor （pifithrin-α, PFT-α） transplantation group （n = 24）. Equal volume of DMEM/F 12, NSC suspension and NSC＋ PFT-α suspension were injected into certain sites of the brain in the above groups 24 h after establishing the model respectively. The expressions of P53 and surviving NSC were detected by immunofluorescence staining 12 h, 24 h and 7 d after transplantation. Results P53 protein was localized in the nuclei of grafted NSC in NSC transplantation group and in the cytoplasm in NSC＋ PFT-α transplantation group. The numbers of surviving cells were （18.20 ± 2.69） cells per high power field in NSC ＋ PFT-α transplantation group and （13.06 ±3.20） cells per high power field in NSC transplantation group. There was a statistical difference in the number of surviving cells between two groups （P 〈 0.05）. Conclusion NSC transplantation combined with P53 inhibitor （PFT-α） have important significance for enhancing the survival rate of grafted NSC.

关 键 词：神经干细胞 细胞移植 基因 p53 脑缺血 模型 动物 大鼠 

分 类 号：R743.31[医药卫生—神经病学与精神病学]