亚低温后脑损伤组织提取液对体外脐带间充质干细胞的影响  

Effect of brain tissue extracts from traumatic brain injury rats treated with mild hypothermia on umbilical cord mesenchymai stem cells in vitro

作　　者：陈以胜 赵明亮[1,2] 梁海乾 陈翀[1,2] 涂悦 张赛[1,2] 孙洪涛 

机构地区：[1]武警部队脑创伤与神经疾病研究所,武警后勤学院附属医院脑科医院 [2]天津市神经创伤修复重点实验室,天津300162

出　　处：《中华创伤杂志》2016年第1期80-84,共5页Chinese Journal of Trauma

基　　金：国家自然科学基金（81471275,81341113,81271392,81401067,81301050）

摘　　要：目的模拟创伤性脑损伤（TBI）脑组织经亚低温治疗后的化学微环境，探讨该环境对体外脐带间充质干细胞（UCMSCs）的影响。方法选取18只SD大鼠按随机数字表法分为3组：假手术组大鼠仅开骨窗，不行冲击损伤；TBI组大鼠仅行冲击损伤；亚低温组大鼠经TBI后行亚低温治疗，各组6只。采用电子脑皮质挫伤撞击仪制作大鼠TBI模型。给予全身亚低温（33％）治疗4h后，获取大鼠模型损伤区域脑组织的匀浆液。模拟脑组织弹性模量制备聚丙烯酰胺水凝胶。分离培养人UCMSCs，将其置于上述水凝胶上进行培养，分别加入各组大鼠的脑组织匀浆液，24h后检测各组细胞的凋亡水平，并更换为UCMSCs的正常培养基。动态观察细胞的生长情况和形态变化，于7d后对其行免疫荧光检测其向神经元分化水平。结果假手术组、TBI组、亚低温组的细胞凋亡率分别为10．42％、73．47％和28．57％，与假手术组比较，TBI组的凋亡细胞明显增多（P〈0．01），而亚低温组可显著逆转TBI组诱导的凋亡细胞增多（P〈0．01）；细胞免疫荧光结果显示，假手术组、TBI组、亚低温组细胞的神经元总体分化率分别为16．48％、2．59％和11．83％，与TBI组比较，亚低温组UCMSCs的神经元分化率明显增高（P〈0．05）。结论TBI后的化学微环境可导致UCMSCs大量凋亡，且向神经元分化的能力大大降低；而亚低温治疗可减少TBI所致UCMSCs的凋亡，并使其分化能力得到一定恢复。Objective To simulate the chemical microenvironment of traumatic brain injury （TBI） under mild hypothermia, and investigate the effect of such microenvironment on umbilical cord mesenchymal stem ceils （ UCMSCs ） in vitro. Methods Eighteen SD rats were allocated to sham- operated group, TBI group and mild hypothermia group according to the random number table, with 6 rats per group. Rat models of TBI were made by electric cortical contusion impactor. After systemic mild hypothermia （33℃） for 4 h, brain tissue homogenate extracts were harvested. Polyacrylamide gels mim- icking the elastic modulus of brain were manufactured. Human UCMSCs were isolated and cultured on the gels, added with brain tissue extracts from each group. After 24 h, the apoptosis level of UCMSCs was checked, and the medium was changed with normal one. Cell growth and morphological changes in each group were given dynamic observation. Seven days later, cell immunoflnorescence was implemented, with the differentiation level of each group estimated. Results Apoptotic rate in TBI group was 73.47%, significantly higher than 10.42% in sham-operated group （P 〈 0.01 ）. While the apoptotic rate was 28. 57% in mild hypothermia group, indicating mild hypothermia significantly reversed the apoptosis of cells in TBI group （ P 〈 0. 01 ）. Cell immunofluorescence demonstrated rate of neuronal differentiation of UCM- SCs in sham-operated group, TBI group and mild hypothermia group was 16.48% , 2.59% and 11.83% respectively. Mild hypothermia resulted in significantly improved neuronal differentiation of UCMSCs after TBI （P 〈 0.05 ）. Conclusions More apoptosis and lower neuronal differentiation ability are observed in UCMSCs in the chemical microenvironment after TBI. However, mild hypothermia significantly reverses the elevation of apoptosis and restores the neuronal differentiation capacity of UCMSCs after TBI.

关 键 词：亚低温 脑损伤 化学微环境 间充质干细胞 组织提取物 

分 类 号：R651.15[医药卫生—外科学]