小鼠小肠上皮细胞经γ射线照射后蛋白质组差异分析  

Differential proteomic analysis of mouse intestinal epithelium irradiated by γ-rays

作　　者：章波[1] 粟永萍[1] 刘晓宏[1] 艾国平[1] 冉新泽[1] 魏永江[1] 王军平[1] 程天民[1] 

机构地区：[1]第三军医大学预防医学系全军复合伤研究所,重庆400038

出　　处：《中华放射医学与防护杂志》2003年第4期234-237,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金资助项目 (3 0 2 3 0 3 60 )

摘　　要：目的　分析γ射线照射后 ,在辐射损伤期和修复期小鼠肠上皮细胞蛋白质组的改变 ,以探讨肠上皮细胞损伤与修复的分子机理。方法　采用 9Gy全身照射BALB c小鼠 ,分别于照射后3h和 72h取小肠制备卷肠切片 ,染色并观察小肠形态。分离未照射小鼠及照射后 3h和 72h小鼠的肠上皮细胞 ,制备蛋白样品 ,采用双向电泳技术分离样品 ,PDQuest软件处理双向电泳图谱 ;并对差异的蛋白质点进行肽质量指纹分析。结果　小鼠肠上皮在照射后 3h以损伤性改变为主 ,72h后出现明显再生修复。采用双向电泳技术在正常对照组、照射后 3h组以及照射后 72h组分别检测到(6 38± 39)、(5 6 6± 32 )和 (5 91± 2 9)个蛋白质点。对其中 2 8个差异蛋白质点进行了肽质量指纹分析 ,经数据库检索 ,初步鉴定为一些与代谢、过氧化应急、信号转导相关的蛋白质。结论　电离辐射能诱导小鼠肠上皮细胞蛋白表达的改变 ,双向电泳技术对从整体方面揭示辐射损伤的分子机理有重要价值。Objective For elucidating the molecular mechanism of reconstruction of intestinal epithelium damaged by ionizing radiation,the proteomes of murine intestinal epithelium from normal and irradiated mice were compared by 2-D electrophoresis. Methods Histopathologic sections of whole small intestine made from \{BALB/c\} mice 3 h and 72 h after total-body irradiation were stained with hematoxylin-eosin.Intestinal epithelial cells were isolated from normal and irradiated mice.The total protein samples prepared by one-step method were used in 2-D electrophoresis,the protein maps were compared and the differential spots were detected with PDQuest analysis software.Twenty-eight different spots were cut off from the gels,digested in gel with trypsin,measured with \{MALDI\}-TOF-MS and searched in databased. Results Small intestinal epithelium was damaged as early as 3 h after irradiation,and reconstructed 72 h later.After Coomassie-staining,the 2-DE image analysis by PDQuest software detected 638±39 protein spots in normal mice group,566±32 spots in 3 hours post irradiation group,and 591±29 spots in 3 days post irradiation group.The 2-DE images showed that proteomes of intestinal epithelium were altered with γ-irradiation.The proteins identified by peptide mass fingerprinting involved in cellular events,including signal transduction,metabolism and oxidative stress responses. Conclusions Gamma-irradiation can induce the protein expression of intestinal epithelium.The technique of 2-D electrophoresis is a useful tool in the study of molecular mechanism of radiation damage.

关 键 词：小鼠 小肠 上皮细胞 Γ射线照射 蛋白质组 差异 分析 双向电泳 肽质量指纹谱 

分 类 号：R811.5[医药卫生—放射医学]