脑创伤后Hes1过表达对成年小鼠海马神经再生的影响  

The effect of Hesl upregulation on neurogenesis after traumatic brain injury

作　　者：李帆[1] 张振 杨新宇 任新亮[1] 

机构地区：[1]长治医学院附属和济医院神经外科,046000 [2]医科大学总医院神经外科,天津300052

出　　处：《中华实验外科杂志》2017年第4期619-621,共3页Chinese Journal of Experimental Surgery

基　　金：山西省自然科学基金面上项目（2014011041-8）

摘　　要：目的 观察腺病毒载体介导的Hes1基因过表达对脑创伤后成年小鼠海马齿状回神经细胞增殖和分化的影响，探讨Hes1过表达对小鼠学习和记忆功能的作用。方法 健康成年雄性C57BL/6小鼠160只，随机分为假手术组、单纯创伤组、阴性对照组和过表达组。构建含小鼠Hes1基因的重组腺病毒，制作小鼠脑创伤液压打击模型。实时荧光定量聚合酶链反应FQ-PCR）检测TBI前1 d和TBI后3 d Hes1 mRNA水平上的表达；Western blot检测TBI前1 d和TBI后3 d Hes1蛋白在海马中组织中的表达，采用溴脱氧尿嘧啶核苷（BrdU）和DCX免疫荧光分别检测脑创伤后3、7 d小鼠海马齿状回神经细胞增殖和分化。脑创伤后7 d和14 d利用水迷宫检测Hes1过表达对小鼠学习和记忆功能的影响。结果 FQ-PCR及Western blot检测显示TBI前1 d和TBI后3 d过表达组海马齿状回组织中Hes1在蛋白和mRNA水平上表达升高（1.87±0.13比0.97±0.13，P＝0.025）；在脑损伤后3 d，过表达组小鼠海马齿状回BrdU表达量较对照组明显降低；脑损伤后7 d过表达组DCX表达量较对照组显著降低。水迷宫检测发现Hes1过表达对成年小鼠逃避潜伏期以及象限百分比无明显影响（P＝0.062）。结论 Hes1基因过表达抑制脑创伤后成年小鼠海马齿状回神经细胞增殖和分化，但对小鼠学习和记忆功能无影响。Objective To investigate the effect of Hes1 upregulation by using an adonoviral vector containing mouse Hes1 gene in adult hippocampal neurogenesis following traumatic brain injury （TBI）, a Morris water maze （MWM） test was employed to investigate the spatial learning and memory capacity of adult mice following injury. Methods One hundred and sixty mice were randomly allocated into four groups： sham-PBS group, sham-PBS-TBI group, Ad5-enhanced green fluorescent protein （EGFP）-TBI group, Ad5-Hes1-TBI group. The TBI model mouse was subjected to lateral fluid percussion injury of （202±2） kPa using a pre-calibrated fluid percussion injury device. Then, Ad5-mRNA and PBS were stereotaetic injected into the hippocampus of the adult C57BL/6 mice and their expressions in the hippocampus were detected. Western blotting and real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） methods were employed to detect the expression level of Hes1 on day 3 following TBI. We performed BrdU or DCX immunofluorescence staining on day 3 or 7 post TBI, to investigate the effect of Hes1 upregulation in adult hippocampal neurogenesis following TBI.Results FQ-PCR and Western blotting analysis suggested that Hes1 was up-regulated in overexpression group at mRNA （1.87±0.13 vs. 0.97±0.13, P=0.025） on day 3 post TBI and protein level; Immunofluorescence staining showed that a significant decrease in the total number of BrdU （＋ ） and DCX （＋ ） cells from the animals with Ad5-mHes1 treated on day 3 or 7 post injury. Morris water maze （MWM） test confirmed that Hes1 overexpression did not lead to a statistically significant change in EL and the target quadrant （P=0.062）.Conclusion These results strongly suggested that increased expression of Hes1 may inhibit adult neurogenesis in DG of hippocampus after TBI and did not lead to a statistically significant change in the spatial learning and memory capacity of adult mice following injury.

关 键 词：Hesl 成年神经再生 脑创伤 MORRIS水迷宫 

分 类 号：R651.15[医药卫生—外科学]