Proteomic Analysis of Glutaraldehyde Fixation on Heterogenic Blood Vessels in Rats
Proteomic Analysis of Glutaraldehyde Fixation on Heterogenic Blood Vessels in Rats
复制标题
戊二醛固定大鼠异种血管的蛋白质组学分析
DOI：
10.1166/jbt.2015.1285
复制
发表时间：
2015
影响因子：
0.1
通讯作者：
Li, Wenbin
中科院分区：
文献类型：
--
作者：
Li, Diankun;Xin, Zhifei;Xu, Xiufang;Li, Wenbin
应用蛋白质组学方法对大鼠体内移植的戊二醛(GA)处理的猪血管的免疫应答进行系统研究。实验分为两组：新鲜组和戊二醛处理组。将20条新鲜和经戊二醛处理的猪肺动脉分别埋藏于大鼠皮下。采用免疫组织化学方法和酶联免疫吸附试验(ELISA)对上述变化进行比较。通过无标记猎枪策略进行蛋白质组学分析。免疫组织化学检测显示，新鲜组第12天、第30天CD4、CD8表达较强。在戊二醛组，它们的表达水平弱于或接近于生鲜组。酶联免疫吸附试验显示IL-6的表达水平与之相似，但IL-2的表达水平在第4天达到峰值。蛋白质组学分析表明，鲜鲜组和戊二醛处理组分别有1278个蛋白质和1064个蛋白质，其中280个蛋白质上调，237个蛋白质下调。这些改变的蛋白质属于三大功能类别：新陈代谢、细胞骨架和应激反应。有6种蛋白的表达存在显著差异，它们与细胞凋亡和免疫/炎症反应有关。戊二醛处理的异种血管引起免疫反应，尽管低于新鲜血管。本研究采用的蛋白质组学鸟枪法能够有效地分离和分析免疫蛋白质，改善了新鲜组和戊二醛处理组的免疫蛋白质表达谱。所鉴定的6种蛋白可能有助于进一步探讨免疫应答机制。
To conduct a systematic study of the immunologic response of rats to transplanted glutaraldehyde (GA)-treated porcine blood vessels in vivo using proteomics. The experiment was divided into two groups: a fresh group and a glutaraldehyde-treated group. Twenty fresh and glutaraldehyde-treated porcine pulmonary arteries were subcutaneously embedded in rats. We compared the changes using immunohistochemistry and enzyme-linked immuno sorbent assay (ELISA). Proteomic analysis via a label-free shotgun strategy was performed. As measured by immunohistochemistry, CD4, CD8 were more strongly expressed on day 12 and day 30 in the fresh group. Their expression levels in the glutaraldehyde group were weaker than or similar to those in the fresh group. From ELISA, the expression levels of Il-6 followed a similar pattern, but IL-2 level reached a peak on day 4. From proteomic analysis, a total of 1278 proteins in fresh group and 1064 proteins in glutaraldehyde-treated group were identified, 280 proteins were up-regulated, and 237 proteins were down-regulated. These altered proteins belonged to three broad functional categories: metabolic, cytoskeletal and stress response. Six proteins to have significant differences in their expression are associated with apoptosis and the immune/inflammatory response. Glutaraldehyde-treated xenogenic blood vessels elicited an immune response, albeit lower than that of fresh blood vessels. The proteomics shotgun strategy used in this study can separate and analyze immunological proteins effectively, which improved the immunological protein expression profiles of fresh and glutaraldehyde-treated groups. The six proteins identified may serve to further explore the immunological response mechanisms.