夜间光暴露小鼠肝脏蛋白质组学分析

王越凡; 张榕; 胡爽; 刘璇; 李鹏翔; 高霞; 魏守刚

doi:10.11847/zgggws1138847

夜间光暴露小鼠肝脏蛋白质组学分析

doi: 10.11847/zgggws1138847

王越凡¹,
张榕¹,
胡爽¹,
刘璇¹,
李鹏翔²,
高霞³,
魏守刚^1, ,

1.
首都医科大学公共卫生学院儿少卫生与妇幼保健学系，北京 100069
2.
杭州市富阳区妇幼保健院
3.
山东省临朐县辛寨中心卫生院卧龙分院

基金项目: 国家自然科学基金（81773451）

详细信息

作者简介:
王越凡（1996 – ），女，内蒙古鄂尔多斯人，硕士在读，研究方向：肥胖的病因机制

通讯作者:
魏守刚，E-mail：shangwei@ccmu.edu.cn

中图分类号: R 575
计量
- 文章访问数: 191
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- PDF下载量: 20
- 被引次数: 0
出版历程
- 接收日期: 2022-04-15
- 网络出版日期: 2023-01-10
- 刊出日期: 2023-03-10

Differential protein expressions in liver of mice exposed to light at night: a proteomics analysis

1.
School of Public Health, Capital Medical University, Beijing 100069, China

摘要

摘要: 目的筛选分析夜间光暴露小鼠肝脏中差异表达蛋白及其生物学意义。方法选择健康雄性C57BL/6J小鼠随机分为夜间光暴露组和对照组，每组8只。夜间光暴露组光照/黑暗周期为24 h/0 h，对照组为12 h/12 h，连续10 d。应用串联质谱标签（TMT）技术寻找并鉴定差异表达蛋白质，对差异蛋白进行GO富集分析和KEGG信号通路分析。结果共鉴定383种差异表达蛋白质，其中215种蛋白质表达水平上调，168种蛋白质表达水平下调。GO功能富集分析和KEGG信号通路分析表明差异表达蛋白质主要分布在胞质部分，分子功能以蛋白结合为主，主要参与蛋白质和脂质等生物代谢过程，作用途径涉及氧化磷酸化信号通路、肿瘤坏死因子（TNF）信号通路、维甲酸诱导基因I（RIG-I）样受体信号通路等。线粒体细胞色素C氧化酶亚基3（MT-CO3）、粒体Fo复合体亚基F2（ATP5J2）、E3泛素连接酶（ITCH）、核因子κB p65亚基（NF-κB p65）、核因子κB激酶抑制剂β亚基（IKBKB）等蛋白分子可能在其中起到关键作用。结论根据差异表达蛋白及其作用分析，夜间光暴露可能有致肥胖、癌症的危险。
- 夜间光暴露 /
- 串联质谱标签(TMT)技术 /
- 蛋白质组学
Abstract: Objective To screen and analyze differentially expressed proteins and their biological significance in the liver of mice exposed to light at night. Methods Totally 16 healthy male C57BL/6J mice were randomly divided into an experimental group (n = 8) exposed to light 24 hours a day continuously for 10 days and a control group (n = 8) with lighting 12 hours a day also for 10 consecutive days. Tandem mass tag (TMT) technology was applied to detect differentially expressed proteins in liver tissues of the mice. The differential protein expressions were analyzed using Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis. Results A total of 383 differentially expressed proteins were identified, of which 215 were expressed at an up-regulated level and 168 proteins were expressed at a down-regulated level. GO enrichment analysis and KEGG signaling pathway analysis revealed that the differentially expressed proteins were mainly distributed in the cytoplasm, and their major molecular functions were protein binding, mainly involved in protein and lipid biological metabolic processes, and their pathways involved in signaling pathways of oxidative phosphorylation, tumor necrosis factor (TNF), and retinoic acid-inducible gene-I (RIG-I)-like receptor. Protein molecules such as mitochondrial cytochrome C oxidase subunit 3 (MT-CO3), mitochondrial Fo complex subunit F2 (ATP5J2), Itchy E3 ubiquitin protein ligase (ITCH), nuclear factor kappa B p65 subunit (NF-κB p65), and inhibitor of nuclear factor Kappa-B kinase subunit Beta (IKBKB) may play a key role. Conclusion In terms of differentially expressed proteins and their functions in liver tissue, light exposure at night may lead to obesity and cancer in mice.
- light exposure at night /
- tandem mass tag technology /
- proteomics

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图 1 差异表达蛋白的火山图

下载: 全尺寸图片幻灯片

表 1 蛋白组学数据库检索参数

检索项目	检索参数
固定修饰（fixed modifications）	carbamidomethyl (C)
可变修饰（variable modifications）	oxidation (M)
酶（enzyme）	trypsin
遗漏酶切位点（maximum missed cleavages）	2
一级质谱误差（peptide mass tolerance）	20 ppm
二级质谱误差（fragment mass tolerance）	0.6 Da
肽段/碎片离子质量数（mass values）	monoisotopic（单同位素）
显著性阈值（significance threshold）	0.05

下载: 导出CSV

表 2 差异表达蛋白GO分类注释

类别	功能分类	差异蛋白比例（%）
生物过程（biological process）	有机氮化合物代谢进程代谢过程大分子定位细胞成分组织定位未知与其他	45 28 6 5 3 13
细胞组分（cell component）	胞质部分膜结合细胞器细胞质其他细胞成分未知与其他	62 14 9 3 12
分子功能（molecular function）	蛋白结合结合多聚腺苷酸RNA结合蛋白催化活性 RNA结合未知与其他	44 15 13 12 2 14

下载: 导出CSV

表 3 KEGG通路富集类别

一级分类	二级分类	代谢通路	涉及差异蛋白数量
代谢	能量代谢	氧化磷酸化	9
	核苷酸代谢	嘧啶代谢	6
	其他氨基酸的代谢	谷胱甘肽代谢	4
	糖类生物合成与代谢	其他类型的O-聚糖生物合成	3
遗传信息处理	翻译	mRNA监测通路	6
环境信息处理	信号转导	TNF信号通路	7
		mTOR信号通路	5
		NF-κB信号通路	6
生物体系统	免疫系统	RIG-I样受体信号通路	5
		NOD样受体信号通路	4
		细胞质DNA感应通路	4
	内分泌系统	脂肪细胞因子信号通路	5
	排泄系统	集合管酸分泌	3
人类疾病	内分泌和代谢性疾病	非酒精性脂肪肝（NAFLD）	8

下载: 导出CSV

表 4 差异蛋白KEGG通路富集结果

代谢通路	上调蛋白质	下调蛋白质
氧化磷酸化（oxidative phosphorylation）	Atp6v1d、Atp5l	Mt-Co3、Atp5j2、Tcirg1、Atp6v0c、Atp5g2、Ndufb8、Cox8a
TNF信号通路（TNF signaling pathway）	Itch、Ripk1、Rela、Tradd、Ikbkb	Icam1、Pgam5
RIG-I样受体信号通路（RIG-I-like receptor signaling pathway）	Ripk1、Isg15、Rela、Tradd、Ikbkb

下载: 导出CSV

参考文献(11)

[1]	Bauer M, Glenn T, Monteith S, et al. The potential influence of LED lighting on mental illness[J]. The World Journal of Biological Psychiatry, 2018, 19(1): 59 – 73. doi: 10.1080/15622975.2017.1417639
[2]	Cho Y, Ryu SH, Lee BR, et al. Effects of artificial light at night on human health: a literature review of observational and experimental studies applied to exposure assessment[J]. Chronobiology Inter-national, 2015, 32(9): 1294 – 1310. doi: 10.3109/07420528.2015.1073158
[3]	Hunter CA, Kartal F, Koc ZC, et al. Mitochondrial oxidative phosphorylation is impaired in TALLYHO mice, a new obesity and type 2 diabetes animal model[J]. The International Journal of Biochemistry and Cell Biology, 2019, 116: 105616. doi: 10.1016/j.biocel.2019.105616
[4]	Moreno-Navarrete JM, Moreno M, Ortega F, et al. CISD1 in association with obesity - associated dysfunctional adipogenesis in human visceral adipose tissue[J]. Obesity, 2016, 24(1): 139 – 147. doi: 10.1002/oby.21334
[5]	Corsi S, Iodice S, Vigna L, et al. Platelet mitochondrial DNA methylation predicts future cardiovascular outcome in adults with overweight and obesity[J]. Clinical Epigenetics, 2020, 12(1): 29. doi: 10.1186/s13148-020-00825-5
[6]	Flaquer A, Baumbach C, Kriebel J, et al. Mitochondrial genetic variants identified to be associated with BMI in adults[J]. PLoS One, 2014, 9(8): e105116. doi: 10.1371/journal.pone.0105116
[7]	Ghnaimawi S, Baum J, Liyanage R, et al. Concurrent EPA and DHA supplementation impairs brown adipogenesis of C2C12 cells[J]. Frontiers in Genetics, 2020, 11: 531. doi: 10.3389/fgene.2020.00531
[8]	Yang XD, Xiang DX, Yang YY. Role of E3 ubiquitin ligases in insulin resistance[J]. Diabetes, Obesity and Metabolism, 2016, 18(8): 747 – 754. doi: 10.1111/dom.12677
[9]	Marino A, Menghini R, Fabrizi M, et al. ITCH deficiency protects from diet - induced obesity[J]. Diabetes, 2014, 63(2): 550 – 561. doi: 10.2337/db13-0802
[10]	Pyo JS, Kang G, Kim DH, et al. Activation of nuclear factor - κB contributes to growth and aggressiveness of papillary thyroid carcinoma[J]. Pathology – Research and Practice, 2013, 209(4): 228 – 232. doi: 10.1016/j.prp.2013.02.004
[11]	Krazinski BE, Kowalczyk AE, Sliwinska-Jewsiewicka A, et al. IKBKB expression in clear cell renal cell carcinoma is associated with tumor grade and patient outcomes[J]. Oncology Reports, 2019, 41(2): 1189 – 1197.