电离辐照损伤小鼠睾丸组织的比较蛋白质组学分析
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摘要
目的:运用生物信息学方法分析电离辐照损伤小鼠的睾丸组织,提取出小鼠辐照组与正常组睾丸组织比较后的差异表达蛋白质,分析其功能和生物学过程。方法:建立小鼠电离辐照损伤模型。运用比较蛋白质组学方法、i TRAQ联合LC-MS/MS检测技术筛选出差异表达蛋白质,进一步利用David、Uniport数据库对差异蛋白进行GO、KEGG富集分析。结果:通过比较小鼠辐照组与正常组睾丸组织共鉴定出蛋白质1633个,包含差异蛋白128个,其中28个表达上调,100个表达下调。GO分析显示差异蛋白主要涉及氧化还原过程、精子发生、ATP代谢等生物学过程。KEGG分析显示主要分布在帕金森氏病、氧化磷酸化、代谢途径等21条信号通路中。结论:利用生物信息学方法对电离辐照损伤小鼠睾丸组织的蛋白质信息进行挖掘,为进一步研究电离辐照对睾丸组织的辐射损伤机制及寻找生物标志物提供参考。
Objective:To investigate the differentially expressed proteins between mouse testicular tissue with ionizing radiation injury and normal testicular tissue using bioinformatics methods and the functions and biological processes of these proteins. Methods:A mouse model of ionizing radiation injury was established. Comparative proteomic methods,isobaric tags for relative and absolute quantitation,and liquid chromatography-tandem mass spectrometry were used to screen out differentially expressed proteins,and then David and Uniport databases were used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genome(KEGG)enrichment analyses of differentially expressed proteins. Results:A total of 1633 proteins were identified in mouse testicular tissue with ionizing radiation injury and normal testicular tissue,with 128 differentially expressed proteins in total,among which 28 were upregulated and100 were downregulated. GO analysis showed that the differentially expressed proteins were mainly involved in biological processes such as oxidation-reduction process,spermatogenesis,and ATP metabolism. KEGG analysis showed that these proteins were mainly distributed in 21 signaling pathways including Parkinson 's disease,oxidative phosphorylation,and metabolic pathways,etc.Conclusion:The bioinformatics method is used for data mining of proteins in mouse testicular tissue with ionizing radiation injury and normal testicular tissue,which provides a reference for further research on the mechanism of ionizing radiation in inducing testicular tissue injury and new biomarkers.
Objective:To investigate the differentially expressed proteins between mouse testicular tissue with ionizing radiation injury and normal testicular tissue using bioinformatics methods and the functions and biological processes of these proteins. Methods:A mouse model of ionizing radiation injury was established. Comparative proteomic methods,isobaric tags for relative and absolute quantitation,and liquid chromatography-tandem mass spectrometry were used to screen out differentially expressed proteins,and then David and Uniport databases were used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genome(KEGG)enrichment analyses of differentially expressed proteins. Results:A total of 1633 proteins were identified in mouse testicular tissue with ionizing radiation injury and normal testicular tissue,with 128 differentially expressed proteins in total,among which 28 were upregulated and100 were downregulated. GO analysis showed that the differentially expressed proteins were mainly involved in biological processes such as oxidation-reduction process,spermatogenesis,and ATP metabolism. KEGG analysis showed that these proteins were mainly distributed in 21 signaling pathways including Parkinson 's disease,oxidative phosphorylation,and metabolic pathways,etc.Conclusion:The bioinformatics method is used for data mining of proteins in mouse testicular tissue with ionizing radiation injury and normal testicular tissue,which provides a reference for further research on the mechanism of ionizing radiation in inducing testicular tissue injury and new biomarkers.
引文
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[20]张科.精子蛋白质AKAP4、GAPDHS和SPANX与A23187诱发顶体反应率在人工授精中的临床应用价值[D].长沙:中南大学,2011.
[21] Jamin A,Wiebe MS. Barrier to Autointegration Factor(BANF1):interwoven roles in nuclear structure,genome integrity,innate immunity,stress responses and progeria[J]. Current Opinion in Cell Biology,2015,34:61-68.
[22] Chiriva-Internati M. Sperm protein 17:clinical relevance of a cancer/testis antigen,from contraception to cancer immunotherapy,and beyond[J]. International Reviews of Immunology,2011,30(2-3):138-149.
[2]刘方娟.小鼠睾丸糖基化蛋白质组学研究[D].南京:南京医科大学,2016.
[3]仵春云,唐开发,张杜平,等.多次低剂量X线照射对小鼠睾丸组织形态及生精细胞P53和bcl-2蛋白表达的影响[J].中国男科学杂志,2013,28(1):17-19.
[4] Fukunaga H,Kaminaga K,Sato T,et al. Application of an ex vivo tissue model to investigate radiobiological effects on spermatogenesis[J].Radiat Res,2018,189(6):661-667.
[5] Xie J,Wang N,Dong X,et al. Graphdiyne nanoparticles with high free radical scavenging activity for radiation protection[J]. ACS Appl Mater Interfaces,2018[Epub ahead of print]. DOI:10.1021/acsami.8b00949.
[6]李敏,赵领,刘真宏,等.川芎提取物对电离辐照损伤实验小鼠脾脏组织细胞凋亡相关蛋白表达的影响[J].中国药理学通报,2018,34(11):1544-1548.
[7]南岩东,杨拴盈,田应选,等.基于鸟枪法蛋白质组学和生物信息学技术对肺鳞癌表达蛋白质谱的分析[J].西安交通大学学报(医学版),2011,32(1):10-16,27.
[8]田鎏,张昌军,刁红录.脂肪酸结合蛋白研究进展[J].生命科学,2017,29(2):164-169.
[9] Kralisch S,Fasshauer M. Adipocyte fatty acid binding protein:a novel adipokine involved in the pathogenesis of metabolic and vascular disease[J]. Diabetologia,2013,56(1):10-21.
[10] Kajimoto K,Takayanagi S,Sasaki S,et al. RNA interference-based silencing reveals the regulatory role of fatty acid-binding protein 4 in the production of IL-6 and vascular endothelial growth factor in 3T3-L1 adipocytes[J]. Endocrinology,2012,153(11):5629-5636.
[11] Ohira H,Fujioka Y,Katagiri C,et al. Butyrate attenuates inflammation and lipolysis generated by the interaction of adipocytes and macrophages[J]. J Atheroscler Thromb,2013,20(5):425-424.
[12] He Y,Li Y,Zhang S,et al. Radicicol,a heat shock protein 90 inhibitor,inhibits differentiation and adipogenesis in 3T3-L1 preadipocytes[J]. Biochemical and Biophysical Research Communications,2013,436(2):169-174.
[13]张丽,皇甫建,肖瑞.脂肪细胞型脂肪酸结合蛋白与2型糖尿病及其相关疾病的研究进展[J].医学综述,2016,22(10):1941-1944.
[14] Obajdin J,Cotter M,Snelling S,et al. Fatty-acid binding protein4(FABP4)as a potential preclinical biomarker of drug-induced kidney injury[J]. Toxicological Sciences,2018,166(2):441-450.
[15] Nakamura M,Michikawa Y,Baba T,et al. Calreticulin is present in the acrosome of spermatids of rat testis[J]. Biochem Biophys Res Commun,1992,186(2):668-673.
[16] Nakamura M,Moriya M,Baba T,et al. An endoplasmic reticulum protein,calreticulin,is transported into the acrosome of rat sperm[J].Experimental Cell Research,1993,205(1):101-110.
[17]王志勇,汤玲珍,高天文. DJ-1抗氧化应激功能研究进展[J].第二军医大学学报,2012,33(1):88-91.
[18]娄钰霞,张钊,王真真,等.帕金森病相关基因DJ-1与氧化应激[J].神经药理学报,2016,6(1):58-64.
[19]周俊豪. CRISP2蛋白在弱精症精液精子中表达下调的分子机制及其临床意义[D].广州:南方医科大学,2015.
[20]张科.精子蛋白质AKAP4、GAPDHS和SPANX与A23187诱发顶体反应率在人工授精中的临床应用价值[D].长沙:中南大学,2011.
[21] Jamin A,Wiebe MS. Barrier to Autointegration Factor(BANF1):interwoven roles in nuclear structure,genome integrity,innate immunity,stress responses and progeria[J]. Current Opinion in Cell Biology,2015,34:61-68.
[22] Chiriva-Internati M. Sperm protein 17:clinical relevance of a cancer/testis antigen,from contraception to cancer immunotherapy,and beyond[J]. International Reviews of Immunology,2011,30(2-3):138-149.
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