SENP3在小鼠脑损伤模型中的表达与细胞内定位  

Expression and cellular distribution of SENP3 in brain tissues after traumatic brain injury in mice

作　　者：玉壮 张顶顶[1] 厉华[1] 闫惠颖[1] 黄立添 张华升 杭春华[1,2,3] 

机构地区：[1]南京大学医学院附属金陵医院神经外科 [2]南方医科大学南京临床医学院 [3]南京军区总医院神经外科,江苏南京210002

出　　处：《中华神经外科疾病研究杂志》2015年第3期196-199,共4页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(81171170);江苏省自然科学基金资助项目(BK2010459);军区医学科技创新课题资助项目(10Z024)

摘　　要：目的研究小泛素类修饰蛋白(SUMO)特异性蛋白酶3(SENP3)在创伤性脑损伤(TBI)脑组织中的分布和表达变化。方法成年雄性ICR小鼠84只,随机分为假手术组和TBI后3 h、6 h、12 h、24 h、3 d、5 d共7组(每组12只)。通过自由落体法制作小鼠TBI模型,应用免疫印迹法(Western blot)法检测TBI后脑组织中SENP3及半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)的蛋白表达变化。应用逆转录聚合酶链反应(RT-PCR)法和免疫组织化学染色法检测TBI后脑组织中SENP3的表达和分布变化。结果 Western blot结果显示,SENP3及Caspase-3蛋白水平在TBI后6 h开始升高,24 h达到高峰。RT-PCR结果显示伤灶周围脑组织中的SENP3的mRNA水平在TBI后逐渐升高,并在12 h达到高峰。免疫组织化学染色检测结果显示假手术组SENP3在细胞核及胞浆内均有表达,但主要位于核内。与假手术组相比,TBI组的脑组织中SENP3阳性细胞的核明显增大、固缩,细胞排列紊乱。结论 SENP3可能参与了TBI后的神经细胞凋亡的机制。Objective The expression and cellular distribution of small ubiquitin-like modifier proteins （SUMO）-specific proteases （SENP3） in brain tissue after traumatic brain injury （TBI） in mice are studied. Methods A total of 84 male ICR （Institute of Cancer Research） mice were randomly divided into seven groups：sham group, TBI 3h, 6 h, 12 h, 24 h, 3 d and 5 d groups, respectively.The brain injury model was induced by free-falling method.The expressions of SENP3 and cysteinyl aspartate specific proteinase 3 （Caspase-3） in the cerebral cortex were analyzed by Western blot.Reverse transcription polymerase chain reaction （ RT-PCR ） assay and immunohistochemical study were performed to evaluate the expression of SENP3 in the brain after TBI.Results The difference between the sham group and 24 h post-TBI group was significant.Western blot indicated that the expression levels of SENP3 and caspase3 proteins were gradually increased from 6 h after TBI and peaked at 24 h.Quantitative real-time PCR demonstrated a gradual increase in SENP3 expression, which peaked at 12 h after TBI and declined subsequently.SENP3-positive cells were observed in both sham and 24 h post-TBI groups.However, robust expression of SENP3 was seldom observed in the sham group, while it was obviously enhanced in the 24 h post-TBI group.Notably, nuclear expression of SENP3 was significantly increased in 24 h post-TBI group.Conclusion SENP3 may participate in the nerve cell apoptosis after TBI.

关 键 词：创伤性脑损伤 SUMO特异性蛋白酶3 免疫组织化学染色 小鼠 

分 类 号：R651[医药卫生—外科学]