早期矽肺损伤的蛋白质组学研究及α晶状体球蛋白B的表达与验证

发布时间：2018-01-16 17:42

本文关键词：早期矽肺损伤的蛋白质组学研究及α晶状体球蛋白B的表达与验证　出处：《中南大学学报(医学版)》2015年08期 　论文类型：期刊论文

【摘要】：目的:建立二氧化硅诱导早期肺损伤大鼠肺组织的二维凝胶电泳图谱,筛选在此过程中起关键作用的靶蛋白分子并进行验证。方法:制作二氧化硅诱导大鼠早期肺损伤模型(实验组),通过HE染色观察肺部形态变化,用双向凝胶电泳法建立二维凝胶图谱,用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析鉴定差异表达蛋白分子,用Western印迹验证差异表达蛋白分子在肺组织中的表达。结果:成功制作二氧化硅早期肺损伤模型,建立实验组及对照组肺组织二维凝胶电泳图谱,二者蛋白表达及分布存在差异,发现差异点40个,选取重复性好,在凝胶上位置和表达适中的共20个进行MALDI-TOF-MS鉴定及数据库搜索并筛选,鉴定出差异蛋白13种,包括α晶状体球蛋白B、间隙蛋白I、肥大细胞蛋白酶等,进一步用Western印迹证实实验组α晶状体球蛋白B表达明显增加,与蛋白质组学结果一致。结论:α晶状体球蛋白B在早期矽肺组织中表达明显升高,提示其可能在矽肺的发生发展中起关键作用。
[Abstract]:Objective: to establish a two-dimensional gel electrophoresis map of lung tissue in rats with early lung injury induced by silica. Methods: early lung injury model of rats induced by silica was made. (experimental group, HE staining was used to observe the changes of lung morphology. Two-dimensional gel map was established by two-dimensional gel electrophoresis. Differential expressed protein was identified by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Expression of differentially expressed protein molecules in lung tissue was verified by Western blotting. Results: early lung injury model of silica was successfully made and two-dimensional gel electrophoresis patterns of lung tissue of experimental group and control group were established. There were differences in protein expression and distribution between the two groups, 40 of them were found to be different, and the selection was reproducible. A total of 20 proteins with moderate position and expression on the gel were identified by MALDI-TOF-MS identification and database search. 13 differential proteins including 伪 -lens globulin B and interstitial protein I were identified. The expression of 伪 -lens globulin B in experimental group was further increased by Western blotting. Conclusion: the expression of 伪 -lens globulin B is significantly increased in early silicosis tissues, suggesting that 伪 -lens globulin B may play a key role in the occurrence and development of silicosis.
【作者单位】：中南大学湘雅三医院病理科;中南大学湘雅三医院耳鼻咽喉头颈外科;中南大学湘雅医院病理科;
【基金】：国家自然科学基金(30170399) 湖南省自然科学基金青年基金(12JJ4079)~~
【分类号】：R135.2
【正文快照】： Methods:Th e animal models of silicosis were established and morphology changes were observedby HE staining,and then the key protein biomarkers in silicosis were identifi ed by 2-DE and matrix-第一作者(First author):初令,Email:catsnow88@126.comassisted l

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