iTRAQ串联质谱筛选尾吊小鼠感染空间诱变大肠杆菌后脾脏差异表达蛋白

发布时间：2018-04-20 07:21

本文选题：iTRAQ + 模拟失重　；参考：《航天医学与医学工程》2017年04期

【摘要】：目的应用同重同位素标记相对与绝对定量技术(iTRAQ),联合液相色谱串联质谱(LC-MS/MS),检测经空间诱变大肠杆菌(T1-13)感染尾吊模拟失重小鼠后,其脾脏组织中的差异蛋白,并探讨其生物学意义。方法 C57BL/6小鼠尾吊模拟失重后以空间诱变大肠杆菌灌胃建立感染模型,采用iTRAQ结合LC-MS/MS技术筛选脾脏组织差异表达蛋白,并对差异蛋白进行GO、pathway富集分析和STRING蛋白互作分析。结果共鉴定出2589个蛋白,尾吊组与对照组相比筛选出378个差异表达蛋白。尾吊染菌后与对照组相比,共筛选出452个差异表达蛋白,STRING蛋白互作网络中有286个差异蛋白间存在相互作用,这些差异蛋白经KEGG分析共得到32条存在显著性的通路,主要为氧化磷酸化通路,代谢通路、蛋白消化和吸收通路、蛋白酶体通路。结论成功筛选出了模拟失重后空间诱变大肠杆菌感染脾脏组织差异表达蛋白,这些差异蛋白可能通过调控代谢通路、不同环境下微生物代谢、颉氨酸/亮氨酸/异亮氨酸降解、溶酶体、碳代谢、吞噬体、丙酸代谢、蛋白酶体参与失重条件下免疫和炎症反应的发生。
[Abstract]:Objective to detect the differential proteins in spleen tissue of mice infected by space mutated Escherichia coli T1-13, using the relative and absolute quantitative techniques of iso-isotopic labeling and absolute quantification, combined with liquid chromatography tandem mass spectrometry (LC-MS / MS), to detect the differential proteins in the spleen of simulated weightlessness mice infected by space mutated Escherichia coli T1-13. Its biological significance was also discussed. Methods after simulated weightlessness in C57BL/6 mice, the infection model was established by space induced Escherichia coli. The differential expression proteins in spleen were screened by iTRAQ and LC-MS/MS technique, and the differential proteins were analyzed by GOA pathway enrichment and STRING protein interaction. Results A total of 2589 proteins were identified and 378 differentially expressed proteins were screened between the tail suspension group and the control group. In comparison with the control group, 286 differentially expressed proteins were found to interact with each other in a total of 452 differentially expressed proteins (STRING) interaction networks, and 32 of the differentially expressed proteins were found to have significant transduction pathways by KEGG analysis. It mainly includes oxidative phosphorylation pathway, metabolic pathway, protein digestion and absorption pathway and proteasome pathway. Conclusion the differentially expressed proteins of splenic tissue induced by simulated weightlessness were successfully screened. These differentially expressed proteins may be mediated by metabolic pathways, microbial metabolism and degradation of antagonic acid / leucine / isoleucine in different environments. Lysosomes, carbon metabolism, phagocytosis, propionic acid metabolism, proteasome participate in immune and inflammatory reaction under weightlessness.
【作者单位】：锦州医科大学;中国人民解放军火箭军总医院;中国航天员科研训练中心航天医学基础与应用国家重点实验室;解放军总医院南楼呼吸科;
【基金】：国家重点基础研究发展(973)计划(2014CB744404)
【分类号】：R852.22

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