SENP3在小鼠脑损伤模型中的表达与细胞内定位

发布时间：2018-02-28 03:51

本文关键词： 创伤性脑损伤 SUMO特异性蛋白酶免疫组织化学染色小鼠　出处：《中华神经外科疾病研究杂志》2015年03期 　论文类型：期刊论文

【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。
[Abstract]:Objective to study the distribution and expression of Ubiquitin modified protein SUMO-specific protease 3 (SENP3) in brain tissue of traumatic brain injury (TBI). Methods 84 adult male ICR mice were used in this study. They were randomly divided into sham operation group and 7 groups (12 rats in each group), and 7 groups (12 rats in each group) were randomly divided into 7 groups: TBI model was established by free falling body method in 3 hours, 6 hours, 12 hours and 24 hours and 5 days after TBI. Western blotting was used to detect the expression of SENP3 and caspase-3 in brain tissue after TBI. Reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical staining were used to detect the expression of SENP3 and caspase-3 in brain tissue. The expression of caspase-3 was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). Results Western blot showed that the levels of SENP3 and Caspase-3 protein began to increase at 6 h after TBI and reached the peak at 24 h. RT-PCR results showed that the mRNA level of SENP3 in the brain tissue around the injured area increased gradually after TBI. The results of immunohistochemical staining showed that SENP3 was expressed in nucleus and cytoplasm of sham-operated group, but mainly in nucleus. Compared with sham-operated group, the nucleus of SENP3 positive cells in brain tissue was significantly increased. Conclusion SENP3 may be involved in the mechanism of neuronal apoptosis after TBI.
【作者单位】：南京大学医学院附属金陵医院神经外科;南方医科大学南京临床医学院;南京军区总医院神经外科;
【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)
【分类号】：R651.15;R-332

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