小鼠脊髓缺血性损伤模型中小泛素化调节子蛋白表达的变化  

The experimental study of small ubiquitin-like modifier conjugation in mice spinal cord ischemia model

作　　者：王正锋[1] 王瑞华[1] 刘献志[1] 翟广[1] 张水军[1] 

机构地区：[1]郑州大学第一附属医院神经外科,450052

出　　处：《中华实验外科杂志》2014年第7期1463-1465,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年基金资助项目（81301008）;河南省教育厅科学技术研究重点资助项目（12A320054）

摘　　要：目的 观察脊髓缺血性损伤是否激活小泛素化调节因子（SUMO）以及SUMO结合蛋白在缺血刺激后的亚细胞定位。方法 制备小鼠脊髓缺血模型,用Western blot法检验脊髓缺血性损伤引起的应激反应,再分别用Western blot和免疫荧光染色法观察缺血脊髓节段中SUMO结合蛋白的表达和亚细胞定位。结果 在脊髓缺血性损伤模型中,应激反应被激活,真核细胞起始因子2（eIF2）、细胞外信号调节激酶（ERK）1/2和蛋白激酶B（Akt）的磷酸化水平明显增高。夹闭8、10、12min组小鼠损伤脊髓节段内SUMO1的表达量分别是对照脊髓节段的（1.15±0.09）、（1.24±0.13）、（1.01±0.99）倍,差异无统计学意义（P〈0.05）,SUMO2/3分别是对照脊髓节段的（5.87±0.31）、（5.40±0.36）、（4.31±0.28）倍,差异有统计学意义（P〈0.01）。免疫荧光染色结果显示损伤脊髓节段SUMO2/3结合蛋白增多并发生明显的核聚集。结论 SUMO2/3在脊髓缺血性损伤的应激反应中起到重要的作用,可能是内源性神经保护途径的关键分子。Objective To determine whether spinal cord ischemia induces small ubiquitin-like modifier （SUMO） conjugation and to observe the subcelluar location of SUMO conjugation. Methods Ac- tivation of stress response pathways after spinal cord ischemia was verified by Western blotting analysis in mice spinal ischemia model. Levels and suhcellular loealizations of SUMO-eonjugated proteins in spinal cords were evaluated by Western blotting analysis and immunol fluorescene staining, respectively. Results Following transient spinal cord ischemia, stress responses were activated as indicated by increased phosphorylation of eukaryotic initiation factor 2 （eIF2）, extracellular signal-regulated kinase （ERK）1/2, and protein kinase B （Akt）. Leves of SUMOl-conjugated proteins were not change significantly after 8, 10, or 12 rain ischemia, 1.15 ±0. 09, 1.24 ± 0. 13, 1.01 ± 0. 99, respectively （P 〉 0. 05）. Levels of SUMO2/3- conjugated proteins were markedly increased： 5.87 ±0. 31,5.40 ±0. 36, 4. 31 ±0. 28, respectively （P 〈 0. 01 ）. The result of immunol fluorescene staining showed that SUMO2/3-conjugation proteins increased markedly and nulei accumulation occurred. Conclusion SUM02/3 maybe a key molecular in internal neuroprotective pathway which played an important role in the stress response of spinal cord ischemia.

关 键 词：脊髓缺血性损伤 小泛素化调节因子 神经保护 

分 类 号：R651.2[医药卫生—外科学]