活性因子对大鼠创伤性脑损伤后内源性神经干细胞增殖和分化的影响  

Effect of active factor on proliferation and differentiation of endogenous neural stem cells after traumatic brain injury in rats

作　　者：任登鹏[1] 李晓红[2] 涂悦[2] 孙洪涛[2] 王景景[2] 王丽娜[2] 陈翀[2] 栾佐[3] 张赛[1] 

机构地区：[1]天津医科大学研究生院,300070 [2]武警后勤学院附属医院脑科医院脑创伤与神经疾病研究所 [3]海军总医院儿科

出　　处：《中华创伤杂志》2016年第9期843-847,共5页Chinese Journal of Trauma

基　　金：国家自然科学基金（81471275,81271392,81401067,81301050）;军队后勤科研计划（CHJ14C022）

摘　　要：目的探讨活性因子对大鼠创伤性脑损伤（TBI）后海马齿状回颗粒下区神经干细胞（NSCs）增殖和分化的影响。方法将SD大鼠按随机数字表法分为假手术组（只暴露硬脑膜，不接受液压冲击）、活性因子组（假手术大鼠给予活性因子治疗）、TBI组[采用液压冲击仪（2．5atm）建立大鼠单侧重型TBI模型]和TBI＋活性因子组（TBI后即刻对侧脑室穿刺并注射活性因子治疗），每组10只大鼠。TBI后7d，40g／L多聚甲醛灌流取材切片或断头取损伤侧海马匀浆，采用免疫荧光染色法检测海马齿状回颗粒下区溴化脱氧尿嘧啶核苷（BrdU）和神经元核抗原（NeuN）的表达，选用连续5张切片进行阳性细胞计数，采用Westernblot检测海马区核转录因子-KB（NF—KB）p65蛋白的表达量。结果与假手术组比较，TBI组伤后7d海马齿状回颗粒下区BrdU＋和NeuN＋细胞数显著增加（P〈0．05）。与TBI组比较，TBI＋活性因子组BrdU＋和NeuN＋细胞明显增加[（207．7±22．5）：（115．9±13．3）]（P〈0．05）。而假手术组和活性因子组BrdU＋和NeuN＋细胞数差异无统计学意义（P〉0．05）。与假手术组比较，TBI组伤后7d海马区NF—KBp65表达增强，而TBI＋活性因子组减弱[（0．81±0．09）：（1．46±0．13）]（P〈0．05），但假手术组和活性因子组海马区NF—KBp65表达无明显变化。结论活性因子可促进大鼠TBI后海马齿状回颗粒下区神经干细胞的增殖和神经元分化，可能与激发神经元生成及降低TBI后海马区NF—KBp65的表达有关。Objective To investigate the effect of active factor on proliferation and differentiation of neural stem cells （NSCs）in the hippocampal subgranular zone after traumatic brain injury（TBI） in rats. Methods SD rats were divided into sham-injured group （only left dura mater exposed）, active factor group （sham injury ＋ active factor therapy）, TBI group （unilateral fluid percussion was used to generate severe TBI）, and TBI ＋ active factor group （TBI ＋ active factor therapy） according to the random number table （n = 10 in each group）. Hippocampal homogenates or brain tissues were harvested at 7 d postinjury. Expressions of BrdU and NeuN in the hippocampal subgranular zone were detected by immunofluorescence staining, and five consecutive sections were used to count BrdU and NeuN positive cells. Level of nuclear factor （NF）-KB p65 in hippocampus was detected by Western blot. Results Number of BrdU and NeuN positive cells in TBI group were greatly higher than that in sham-injured group （P 〈 0.05 ）. Number of BrdU and NeuN positive cells in TBI ＋ active factor group was greatly higher than that in TBI group [ （ 207.7±22.5 ） vs. （ 115.9±13.3 ） ] （ P 〈 O. 05 ）. There were no significant differences in BrdU and NeuN positive cells between sham-injured group and active factor group （ P 〉 0.05 ）. NF-KBp65 presented over-expression in TBI group （0.81±0. 09 ） while reversed in TBI ＋ active factor group （1.46±0. 13）, when compared to sham-injured group （P 〈0. 05）. However, no significant change in NF-KBp65 was noted between sham-injured group and active factor group （P 〉0. 05）. Conclusion Active factor therapy can promote proliferation and neuronal differentiation of NSCs in the hippocampal subgranular zone after TBI in rats and the possible mechanism may relate to activation of the NSCs and inhibition of over-expression of NF-KBp65.

关 键 词：颅脑创伤 神经干细胞 活性因子 

分 类 号：R651.15[医药卫生—外科学]