脂肪间充质干细胞分泌组对颅脑创伤后大鼠神经功能的影响  

Effect of adipose-derived mesenchymal stem cells secretome on neurological function of rats after traumatic brain injury

作　　者：徐超[1] 李晓红[1] 王晶[1] 史洪建 王景景[1] 程军 陈江龙[1] 孙洪涛[1] 涂悦[1] 张赛[1] Xu Chao;Li Xiaohong;Wang Jing;Shi Hongjian;Wang Jingjing;Cheng Jun;Chen Jianglong;Sun Hongtao;Tu Yue;Zhang Sai(Institution of Brain Trauma and Neurology Disease of Affiliated Hospital of Logistics University of PAl）,Tianjin 300162,China;Department of MRI of Affiliated Hospital of Logistics University of PAP,Tianjin 300162,China)

机构地区：[1]武警后勤学院附属医院颅脑创伤与神经修复研究所,天津300162 [2]武警后勤学院附属医院核磁共振科,天津300162

出　　处：《遵义医学院学报》2018年第4期418-423,430,共7页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:81401067;81771352;81471275;81671222;81771350);天津市自然科学基金资助项目(NO:15JCQNJC11100;15JCYBJC28100)

摘　　要：目的研究脂肪间充质干细胞分泌组(Adipose-derived mesenchymal stem cells secretome,ASC-ST)对大鼠颅脑创伤(Traumatic brain injury,TBI)后神经功能的影响及其潜在的机制。方法将45只雄性SD大鼠随机分为3组,假手术组(Sham组)、单纯TBI组(TBI组)和TBI后静脉注射ASC-ST组(TBI+ST组),每组15只。Sham组只开骨窗,TBI组大鼠在eCCI打击后每日经尾静脉注射0.3 mL生理盐水,TBI+ST组在打击后相同时间点经尾静脉注射0.2 mL ASC-ST和0.1 mL生理盐水,连续注射7 d。在TBI后10～14 d对各组大鼠进行水迷宫实验检测高级神经功能;在TBI后7 d和14 d对TBI和TBI+ST组大鼠行头颅MRI,DTI扫描,测量FA值评估神经纤维损伤;检测损伤周围皮层中Bcl-2和Bax蛋白的含量;并在TBI后14 d用免疫荧光检测损伤周围皮层NeuN^+神经元的含量。结果在TBI后,TBI和TBI+ST组大鼠寻找平台的潜伏期显著高于Sham组(P<0.05),在TBI后14 d,TBI+ST组大鼠的潜伏期显著低于TBI组,当去掉平台后,其在60 s内穿过平台位置的次数显著高于TBI组(P<0.05),但与Sham组无显著差异。在TBI后7 d,TBI+ST组大鼠损伤周围皮层FA值较TBI组高,Bcl-2的含量显著高于TBI组、Bax的含量则低于TBI组,但Bcl-2和Bax的含量均显著高于Sham组。而在TBI后14 d,TBI+ST组大鼠损伤周围皮层FA值与TBI组无显著差异,但损伤对侧皮层、双侧胼胝体的FA值均高于TBI组(P<0.05);损伤周围皮层只有Bax的含量显著低于TBI组(P<0.05),且与Sham组无显著差异。同时,在TBI后14 d,TBI+ST组大鼠损伤周围皮层中NeuN+细胞显著多于TBI组(P<0.05)。结论 ASC-ST能够通过减少神经细胞坏死凋亡及减轻神经纤维损伤改善大鼠TBI后的神经功能预后。Objective To evaluate the effect of adipose-derived mesenchymal stem cells secretome（ASC-ST） exerted on neurological function of rats after traumatic brain injury（TBI） and their potential mechanisms. Methods 45 male Sprague-Dawley rats were randomly divided into 3 groups：Sham surgery（Sham）,TBI and saline injection（TBI）,and TBI and ASC-ST injection（TBI ＋ ST） groups. Each group contained 15 rats. Rats from Sham group only received craniotomy. Rats in the TBI group were injected with 0. 3 ml of normal saline by tail vein after the eCCI attack while the TBI ＋ ST group rats were injected 0. 2 ml ASC-ST and 0. 1 ml normal saline at the same time point after TBI. The treatments were persisted continuously for 7 days. The spatial cognition and memory were measured with Morris water maze test from 10 to 14 days after TBI. The damage of nerve fibers was estimated by DTI scanning on 7 and 14 days after TBI. The levels of Bcl-2 and Bax in the injured cortex were measured by western-blot. Meanwhile,the number of Neu N positive neurons in the injured cortex was detected by immunofluorescence on 14 days after TBI. Results The latency of TBI and TBI ＋ ST rats was significantly higher than that of Sham group（P〈0. 05）. The latency of rats from TBI ＋ ST group was significantly lower than that of TBI group rats on 14 days after TBI（P〈0. 05）. After the platform was removed on 14 days after TBI,rats in TBI ＋ ST groups passed through this position more times than TBI group in 60 s,but there was no significant difference between TBI ＋ ST and Sham groups（P〈0. 05）. On 7 days after TBI,TBI ＋ ST group rats got higher FA values in ipsilateral cortex than TBI group rats. However,there was no significant difference in FA values of between TBI ＋ ST and TBI groups in other parts. Meanwhile,level of Bcl-2 in TBI ＋ ST group was higher than that in TBI group,and the level of Bax was lower than that in TBI group. The contents of Bcl-2 and Bax were higher than in the Sham group. On 14 days aft

关 键 词：颅脑创伤 间充质干细胞 分泌组 神经功能 神经纤维 

分 类 号：R651.15[医药卫生—外科学]