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成年小鼠脑创伤后海马组织中Hes1的表达

张奇1,郇林春1,赵旺淼1,赵杰2,杨树源1,杨新宇1   

  1. 1. 天津医科大学总医院
    2. 天津市人民医院
  • 收稿日期:2010-07-08 修回日期:2010-07-26 出版日期:2010-12-15 发布日期:2010-12-15
  • 通讯作者: 张奇

Analysis of the expression of Hes1 in adult mouse hippocampus after traumatic brain injury

  • Received:2010-07-08 Revised:2010-07-26 Published:2010-12-15 Online:2010-12-15
  • Contact: zhang qi

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2011

摘要：

目的:观察和分析创伤性脑损伤后成年小鼠海马组织中Hes1的表达情况。方法:雄性C57BL/6小鼠66只,随机分为假手术(NC)组33只和创伤性脑创伤(TBI)组33只,通过单侧液压性损伤(FPI)的方法建立小鼠创伤性脑创伤模型。分别在成模后6h、1d、3d和7d通过颈椎脱臼处死小鼠,2组每个时间点各选取小鼠6只,采用实时荧光定量PCR检测海马中Hes1 mRNA的表达;创伤后3d,每组分别选取小鼠6只和3只分别用Western blot法和荧光免疫组织化学染色方法检测Hes1蛋白的表达情况。实时荧光定量PCR和Western blot均以GAPDH作为内参照。结果:TBI组所有标本中均检测到Hes1的表达。TBI组与相应的NC组海马组织中Hes1 mRNA的表达情况各时点比较差异均有统计学意义(P<0.05);TBI组创伤后3d海马的蛋白表达量与NC组比较差异均有统计学意义(P<0.05)。Hes1蛋白在NC组小鼠伤侧海马组织的细胞中广泛表达,而TBI组创伤后3d Hes1蛋白的表达量明显降低。结论:Hes1可能参与了神经发生过程,并在随后的损伤修复中起重要的作用。

关键词: Hes1, 海马, FPI, 创伤性脑损伤, 神经干细胞

Abstract: Objective:To observe and analyze the changes of the expression level of Hes1 in adult mouse hippocampus after traumatic brain injury(TBI). Methods:The C57BL/6 mice were randomly divided into sham operation group(NC,n=33) and traumatic brain injury group(TBI,n=33). Here,we established the experimental TBI model by means of lateral fluid percussion injury(FPI).Then，mice were killed by cervical dislocation 6h,1d,3d and 7d after operation. The expression of Hes1 gene in adult mouse hippocampus was detected by real time PCR and Western blot ,fluorescence immunohistochemistry staining technique at mRNA and protein level.GAPDH is as the internal reference in real time PCR and Western blot. Results:Hes1 was expressed in all hippocampus of TBI group mice. Compared with the NC group（1.03±0.19，1.02±0.14，1.04±0.16，1.06±0.20）, the expression of Hes1/GAPDH mRNA in traumatic lateral hippocampus 6 h , 1d, 3d and 7d after TBI were respectively 6h(1.37±0.14),1d(0.65±0.17),3d(0.46±0.14)和7d(0.82±0.10); the expression of Hes1 protein in traumatic lateral hippocampus on 3d after TBI was (1.25±0.21) contrast to NC groups’ (0.39±0.17).The differences is statistically significant(P<0.05). Fluorescence immunohistochemistry staining shows that Hes1 protein expression was wide in the hippocampus cells of NC group mice, compared with which, TBI group mice’ 3 days after injury was significantly decreased. Conclusion: Hes1 maybe involved in the neurogenesis process and plays an important role in subsequent repair.

Key words: Hes1, Hippocampus, FPI, traumatic brain injury, Neural stem cell