| 章波,粟永萍,刘晓宏,等.小鼠小肠上皮细胞经γ射线照射后蛋白质组差异分析[J].中华放射医学与防护杂志,2003,23(4):234-237.ZHANG Bo,SU Yongping,LIU Xiaohong,et al.Differential proteomic analysis of mouse intestinal epithelium irradiated by γ-rays[J].Chin J Radiol Med Prot,2003,23(4):234-237 |
| 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 |
| 投稿时间：2003-01-30 |
| DOI： |
| 中文关键词: 电离辐射 肠上皮细胞 双向电泳 肽质量指纹谱 |
| 英文关键词:Ionizing radiation Intestinal epithelial cells 2-D electrophoresis Peptide mass fingerprinting |
| 基金项目:国家自然科学基金资助项目(30230360) |
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| 中文摘要: |
| 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(638±39)、(566±32)和(591±29)个蛋白质点。对其中28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。 |
| 英文摘要: |
| 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. |
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