外泌体蛋白质组学分析白念珠菌感染小鼠的标志物

doi:10.16139/j.1007-9610.2021.03.018

摘要/Abstract

摘要：

目的：本研究通过血清外泌体蛋白质组学分析,寻找侵袭性白念珠菌感染的标志物。方法：尾静脉注射白念珠菌菌液建立小鼠模型。提取血清外泌体,分离富集血清外泌体,液相色谱-质谱分析外泌体蛋白质组学。构建金黄色葡萄球菌和大肠埃希菌感染模型,比较外泌体差异蛋白在3个感染组的表达差异。结果：共鉴定4 055个不同肽段,对应418个蛋白质。以符合白念珠菌组/正常对照组差异变化<0.5倍至>2.0倍,同时P<0.05为标准,筛查出差异蛋白10个。对与白念珠菌感染密切相关的蛋白质Cathepsin B、Pentraxin 3进行ELISA鉴定,建立受试者工作特征曲线诊断模型,发现外泌体差异蛋白具有很高的灵敏度和特异度。在3个不同的感染组中,发现外泌体差异蛋白大量在白念珠菌感染组中表达,在非白念珠菌感染组中少量表达。结论：Cathepsin B、Pentraxin 3与白念珠菌感染密切相关,可作为白念珠菌感染的分子标志。

关键词: 蛋白质组学, 生物标志物, 外泌体, 白念珠菌感染

Abstract:

Objective To analyze serum exosome using proteomic in mice with Candida albicans infection and to find out marker. Methods Mice model of Candida albicans infection was established by injection of Candida albicans liquid through tail vein. Exosome in serum were extracted and analyzed by liquid chromatography-mass spectrometry/mass spectrometry. Mice models of both Staphylococcus aureus infection and Escherichia coli infection were established to compare the protein expression in exosome among three groups of infection. Results A total of 4 055 peptides and 418 proteins were identified. With the ratio <0.5 or >2.0 times between Candida albicans infection group and normal control group and with P<0.05, 10 of different proteins were screened out. Both Cathepsin B and Pentraxin 3 which were related with Candida albicans infection were identified by ELISA. Receiver operating characteristic curves were used to evaluate proteins and the different proteins in exosome were found to have high sensitivity and specificity. In 3 groups of Candida albicans infection, Staphylococcus aureus infection and Escherichia coli infection, it was found that exosome proteins expressed more in Candida albicans infection group and less in the other two infection group. Conclusions Proteins including Cathepsin B and Pentraxin 3 were related to Candida albicans infection, which could be used as molecular markers of Candida albicans infection.

Key words: Proteomics, Biomarker, Exosome, Candida albicans infection

中图分类号:

R519.3

王泽田, 吴春荣, 齐越, 徐丹, 孙科远, 唐建国. 外泌体蛋白质组学分析白念珠菌感染小鼠的标志物[J]. 外科理论与实践, 2021, 26(03): 262-268.

WANG Zetian, WU Chunrong, QI Yue, XU Dan, SUN Keyuan, TANG Jianguo. Proteomics analysis of exosome in mice with Candida albicans infection for biomarker[J]. Journal of Surgery Concepts & Practice, 2021, 26(03): 262-268.

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参考文献 25

[1]	扈东营, 姜伟伟, 李航, 等. 侵袭性念珠菌感染流行病学现状和治疗进展[J]. 世界临床药物, 2020, 41(5):5-10.
[2]	Liao Y, Chen M, Hartmann T, et al. Epidemiology of opportunistic invasive fungal infections in China: review of literature[J]. Chin Med J (Engl), 2013, 126(2):561-568.
[3]	Wortzel I, Dror S, Kenific CM, et al. Exosome-mediated metastasis: communication from a distance[J]. Cell Press, 2019, 49(3):347-360.
[4]	Ghamloush F, Ghayad SE, Rammal G, et al. The PAX3-FOXO1 oncogene alters exosome miRNA content and leads to paracrine effects mediated by exosomal miR-486[J]. Scientific Reports, 2019, 9(1):14242. doi: 10.1038/s41598-019-50592-4 pmid: 31578374
[5]	Zhang J, Li S, Li L, et al. Exosome and exosomal microRNA: trafficking, sorting, and function[J]. Genomics Proteomics Bioinformatics, 2015, 13(1):17-24. doi: 10.1016/j.gpb.2015.02.001 URL
[6]	童译庆, 王秋月, 李康馨, 等. 早期白假丝酵母血流感染大鼠模型的血浆代谢组学分析[J]. 微生物与感染, 2019, 14(1):30-38.
[7]	吴丹, 周树生, 胡仕静, 等. NLRP3炎性小体在金黄色葡萄球菌血流感染小鼠免疫反应中的作用[J]. 中国感染与化疗杂志, 2016, 16(4):411-418.
[8]	杨明, 麻雅婷, 何赏, 等. 白细胞介素-6和白细胞介素-10在不同细菌性血流感染小鼠模型中的表达及意义[J]. 感染、炎症、修复, 2017, 18(1):18-22.
[9]	严蕾, 吴春荣, 王晨, 等. 肠缺血再灌注损伤后白色念珠菌深部侵袭研究[J]. 临床急诊杂志, 2019, 20(4):39-44.
[10]	Walker LA, Maccallum DM, Bertram G, et al. Genome-wide analysis of Candida albicans gene expression patterns during infection of the mammalian kidney[J]. Fungal Genet Biol, 2009, 46(2):210-219. doi: 10.1016/j.fgb.2008.10.012 URL
[11]	Kobayashi-Sakamoto M, Tamai R, Isogai E, et al. Gastrointestinal colonisation and systemic spread of Candida albicans in mice treated with antibiotics and prednisolone[J]. Microbial Pathogenesis, 2018, 117:191-199. doi: S0882-4010(17)30878-1 pmid: 29477742
[12]	Bhringer M, Pohlers S, Schulze S, et al. Candida albicans infection leads to barrier breakdown and a MAPK/NF-κB mediated stress response in the intestinal epithelial cell line C2BBe1[J]. Cell Microbiol, 2016, 18(7):889-904. doi: 10.1111/cmi.12566 URL
[13]	Lei Y, Yang C, Tang J. Disruption of the intestinal mucosal barrier in Candida albicans infections[J]. Microbiol Res, 2013, 168(7):389-395. doi: 10.1016/j.micres.2013.02.008 pmid: 23545353
[14]	Salehi M, Sharifi M. Exosomal miRNAs as novel cancer biomarkers: challenges and opportunities[J]. J Cell Phy-siol, 2018, 233(9):6370-6380.
[15]	Yan B, Zhang Y, Liang C, et al. Stem cell-derived exosomes prevent pyroptosis and repair ischemic muscle injury through a novel exosome/circHIPK3/ FOXO3a pathway[J]. Theranostics, 2020, 10(15):6728-6742. doi: 10.7150/thno.42259 URL
[16]	Jia Y, Chen Y, Wang Q, et al. Exosome: emerging biomarker in breast cancer[J]. Oncotarget, 2017, 8(25):41717-41733. doi: 10.18632/oncotarget.16684 URL
[17]	Kourembanas S. Exosomes: vehicles of intercellular signaling, biomarkers, and vectors of cell therapy[J]. Annu Rev Physiol, 2015, 77:13-27. doi: 10.1146/annurev-physiol-021014-071641 pmid: 25293529
[18]	Lai RC, Yeo RW, Tan KH, et al. Exosomes for drug delivery-a novel application for the mesenchymal stem cell[J]. Biotechnol Adv, 2013, 31(5):543-551. doi: 10.1016/j.biotechadv.2012.08.008 URL
[19]	黄媛, 王艺磊, 冯建军, 等. 组织蛋白酶B在水生动物中的研究进展[J]. 生命科学, 2016, 28(11):1384-1390.
[20]	Wang Y, Jia L, Shen J, et al. Cathepsin B aggravates coxsackievirus B3-induced myocarditis through activating the inflammasome and promoting pyroptosis[J]. PLoS Pathog, 2018, 14(1):e1006872. doi: 10.1371/journal.ppat.1006872 URL
[21]	于波, 李世拥, 安萍, 等. 组织蛋白酶B在结直肠癌中的表达及临床意义[J]. 中华胃肠外科杂志, 2005, 8(6):507-509.
[22]	蔡胜, 李振英. Pentraxin 3与补体的相互作用在系统性红斑狼疮中研究进展[J]. 世界最新医学信息文摘, 2019, 19(98):153-155.
[23]	He X, Han B, Liu M. Long pentraxin 3 in pulmonary infection and acute lung injury[J]. Am J Physiol Lung Cell Mol Physiol, 2007, 292(5):L1039-L1049.
[24]	马伟立. 血清外泌体膜蛋白GPC-1在乳腺癌诊断中的应用价值探讨[D]. 河北医科大学, 2018,35-36.
[25]	曹佳实. 基于骨巨细胞瘤患者血清外泌体的蛋白质组学研究[D]. 中国人民解放军海军军医大学, 2019,32-33.

蛋白质 ID	蛋白质名称	基因名称	肽段数	比值（白念珠菌组/正常对照组）	P值
D3ZT94	Pentraxin 3	Ptx3	9	8.546177	2.25E-05
H6X2X0	reactive protein	Crp	7	2.333257	0.000325
A0A0H2UHI5	Serine protease inhibitor A3N	Serpina3n	28	4.053251	2.71E-05
P20059	Hemopexin	Hpx	27	2.442018	0.000254
Q5BKC4	Complement component C9	C9	28	2.748338	4.88E-05
Q6IN22	Cathepsin B	Ctsb	3	3.425430	0.034993
P27139	Carbonic anhydrase 2	Ca2	6	8.901092	4.17E-06
P06238	Alpha-2-macroglobulin	A2m	82	11.166290	5.72E-06
A0A0H2UHF8	Alpha-1-acid glycoprotein	Orm1	7	16.030460	5.07E-10
Q7TQ70	Fibrinogen alpha chain	Fga	26	3.043354	4.64E-05