60Co γ射线全身照射大鼠尿液的非标记定量蛋白质组学分析

doi:10.3969/j.issn.1004-616x.2021.03.003

癌变·畸变·突变 ›› 2021, Vol. 33 ›› Issue (3): 172-177,192.doi: 10.3969/j.issn.1004-616x.2021.03.003

⁶⁰Co γ射线全身照射大鼠尿液的非标记定量蛋白质组学分析

向嘉琪¹, 孙佳丽^1,2, 王成芳¹, 刘青杰¹, 田梅¹

1. 中国疾病预防控制中心辐射防护与核安全医学所, 辐射防护与核应急中国疾病预防控制中心重点实验室, 北京 100088;
2. 北京市通州区疾病预防控制中心, 北京 101100

收稿日期:2020-12-30 修回日期:2021-04-20 出版日期:2021-05-30 发布日期:2021-06-09
通讯作者: 田梅，E-mail：tianmei@nirp.chinacdc.cn E-mail:tianmei@nirp.chinacdc.cn
作者简介:向嘉琪，E-mail：xiangjq1995@163.com。

Analyses of label-free quantitative proteomics on rat urine after ⁶⁰Co γ ray irradiation

XIANG Jiaqi¹, SUN Jiali^1,2, WANG Chengfang¹, LIU Qingjie¹, TIAN Mei¹

1. China CDC Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088;
2. Beijing Tongzhou Center for Disease Control and Prevention, Beijing 101100, China

Received:2020-12-30 Revised:2021-04-20 Online:2021-05-30 Published:2021-06-09

摘要/Abstract

摘要： 目的：鉴定全身照射大鼠尿液中的异常表达蛋白质，筛选辐射敏感生物标志，并分析其功能和相关生物学过程。方法：以未照射大鼠为阴性对照，分别采用1、5 Gy （剂量率1 Gy/min）的⁶⁰Co γ射线全身照射30只大鼠，收集照射后第1和第3天的尿液样本，采用非标记（label-free）质谱技术检测样本中所鉴定蛋白的相对表达水平，筛选差异表达蛋白，进一步使用Uniport、DAVID数据库对其进行基因本体（GO）分析、京都基因与基因组百科全书（KEGG）富集分析，并使用STRING在线网站构建蛋白-蛋白相互作用网络。结果：本研究各样本共鉴定出1 069种蛋白。照射后第1和第3天，与对照组比较1和5 Gy照射组均出现差异表达的蛋白279种，其中190种表达上调，97种表达下调。GO富集分析结果显示差异蛋白主要涉及对药物反应、氧化还原、老化等生物学过程，分布在细胞外泌体、细胞外空间等区域，发挥蛋白质同源二聚化活性、蛋白质结合以及钙离子结合等分子功能。KEGG富集分析结果显示差异蛋白主要涉及抗生素生物合成、谷胱甘肽代谢、碳代谢、代谢途径等多种信号通路。结论：γ射线照射后大鼠尿液蛋白质组学的综合分析表明，Gusb、Reg3g、GCLM、AZGP1蛋白上调和Ly6d蛋白下调可作为辐射敏感候选生物标志。

关键词: 电离辐射, 非标记定量蛋白质组学, 质谱, 液相色谱-质谱/质谱

Abstract: OBJECTIVE： To identify and analzye differentially-expressed and radiation-sensitive protein biomarkers in urines from rats after their total- body irradiation. METHODS： 30 rats were irradiated whole body with single doses of 0，1，5 Gy ⁶⁰Co γ rays at a dose rate of 1 Gy/min. Urine samples were collected at 1 and 3 days post irradiation. A label- free mass spectrometric technology was used to measure relative levels of differentially expressed proteins. Uniport and DAVID databases were conducted using Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The STRING online website was used to construct a protein-protein interaction network. RESULTS： A total of 1 069 proteins were identified. Compared with the negative control group， a total of 279 shared proteins were differentiallyexpressed in the 1 and 5 Gy irradiation groups after irradiation at 1 and 3 days， of which 190 were upregulated and 97 were down-regulated. Results from the GO analyses show that the differentially-expressed proteins were mainly involved in the biological processes such as response to drug， oxidation-reduction process，and aging. Most of the cellular components of these screened proteins were extracellular exosomes and extracellular space，and these proteins exerted protein homodimerization activity，protein binding and calcium ion binding and other molecular functions. KEGG enrichment analyses indicate that changes of the differentiallyexpressed proteins mainly participated in several signal pathways including antibiotic biosynthesis， glutathione metabolism， carbon metabolism， and metabolic pathways. CONCLUSION： Our proteomic study is a comprehensive analysis of the rat urine proteome following radiation exposure. Our analyses suggest that the up-regulated proteins Gusb，Reg3g，GCLM，AZGP1 and down-regulated protein Ly6d can be used as candidate biomarkers of radiation sensitivity for future research.

Key words: ionizing radiation, label-free quantitative proteomics, mass spectrometry, LC-MS/MS

中图分类号:

R994.6

向嘉琪, 孙佳丽, 王成芳, 刘青杰, 田梅. ⁶⁰Co γ射线全身照射大鼠尿液的非标记定量蛋白质组学分析[J]. 癌变·畸变·突变, 2021, 33(3): 172-177,192.

XIANG Jiaqi, SUN Jiali, WANG Chengfang, LIU Qingjie, TIAN Mei. Analyses of label-free quantitative proteomics on rat urine after ⁶⁰Co γ ray irradiation[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(3): 172-177,192.

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⁶⁰Co γ射线全身照射大鼠尿液的非标记定量蛋白质组学分析

Analyses of label-free quantitative proteomics on rat urine after ⁶⁰Co γ ray irradiation

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