预热适应成纤维细胞NIH-3T3应激保护相关蛋白的初步筛选  

Primary screening of stress protective proteins in heat-preconditioned fibroblast NIH-3T3

作　　者：陈雪梅[1] 邹飞[1] 

机构地区：[1]南方医科大学劳动与职业卫生学系,广东广州510515

出　　处：《中国病理生理杂志》2006年第8期1493-1496,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30371575No.30500580);广东省自然科学基金资助项目(No.5300465)

摘　　要：目的:初步分析应激与适应对蛋白质合成的影响,筛选应激适应相关蛋白。方法:通过双向电泳分离总蛋白、PDQUEST软件分析、肽质量指纹图谱分析候选蛋白肽段组成,分析应激与适应对细胞蛋白质合成的影响。初步鉴定应激适应蛋白质点的类别。结果:预适应后再次热应激组表达增加的蛋白质点在各分子量范围均有分布,而直接热应激组表达增加的蛋白质点大多局限于低分子量范围。肽质量指纹图谱分析候选蛋白序列,初步鉴定了Tubu linβ、中间丝vim entin、起始因子eIF-4AI、Eno1蛋白与应激适应密切相关。结论:细胞在紧急应激状态,可能会倾向于选择性地合成能够迅速翻译的小分子量蛋白质,应对应激损伤。预适应可通过提高细胞内保护性蛋白的贮备量,在再次应激时发挥保护作用。Tubu linβ、中间丝vim entin、起始因子eIF-4AI、Eno1蛋白差异表达,提示细胞骨架、蛋白质合成、糖代谢通路在应激适应中的重要作用。AIM： To establish stress adaptation model of mouse fibroblast cell line NIH -3T3 to analyze the effect of stress and adaptation on protein synthesis, and to screen for stress adaptation related proteins. METHODS： A stress - adapted cell model was established by thermal preconditioning （42℃, 20 min）. Total cytolytes were separated by 2 - DE, analyzed by PDQUEST software, and the selected differential expression spots were detected by MOLDI - TOF. The effect of stress and adaptation on protein synthesis was studied. The stress adaptation related spots were identified by PMF. RESULTS ： Comparative proteomics method by 2 - DE was used to find different distributions of total proteins of heat stress group and thermal preconditioning group. Expression - increased protein spots were found almost limited in low molecular weight range in directly stress group, whereas expression - increased protein spots in thermal preconditioning group have more extensive molecular weight distribution. PMF results showed that tubulinβ, vimentin, eIF -4AI, Enol protein might be related to stress adaptation. CONCLUSION： 2 - DE analysis suggested, cell might favor to synthesize low small molecular weight protein to deal with hostile stress. Cellular protein storage might be increased by preconditioning, and may play a protection role during successive stress. The increased expression of tubulinβ, vimentin, eIF- 4AI, Enol protein suggests that cytoskeletons, protein synthesis pathway and glycometabolism pathway may play an important role in stress adaption.

关 键 词：热预处理 应激 蛋白质组 成纤维细胞 

分 类 号：R363[医药卫生—病理学]