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

来源 :航天医学与医学工程 | 被引量 : 0次 | 上传用户:yokuchan
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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。 OBJECTIVE: To study the effects of space-induced E.coli (T1-13) infection on the tail-suspended mice of simulated weightlessness by iTRAQ and LC-MS / MS, Its spleen and its differential proteins, and to explore its biological significance. Methods C57BL / 6 mice were subjected to tail-suspension simulation to establish an infection model by space-mutated E. coli. The differentially expressed proteins of spleen tissues were screened by iTRAQ and LC-MS / MS, and the differentially expressed proteins were analyzed by GO and pathway Interaction with STRING protein. Results A total of 2589 proteins were identified, and 378 differentially expressed proteins were screened from the tail suspension group compared with the control group. Compared with the control group, a total of 452 differentially expressed proteins were screened out from the tail-trapping bacterium. There were 286 differential proteins in the interaction network of STRING proteins. There were 32 significant differences in these differential proteins analyzed by KEGG Pathways, mainly oxidative phosphorylation pathway, metabolic pathway, protein digestion and absorption pathways, proteasome pathway. Conclusion The differentially expressed proteins in spleen of Escherichia coli infected with space-mutated Escherichia coli after simulated-weightlessness were successfully screened. These differential proteins may be regulated by metabolic pathway, microbial metabolism in different environments, degradation of valine / leucine / isoleucine, Enzymes, carbon metabolism, phagosome, propionate metabolism, proteasome involved in the immune and inflammatory reactions under the condition of weightlessness.
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