您的位置:山东大学 -> 科技期刊社 -> 《山东大学学报(医学版)》

山东大学学报(医学版) ›› 2015, Vol. 53 ›› Issue (3): 12-16.doi: 10.6040/j.issn.1671-7554.0.2014.766

• 基础医学 • 上一篇    下一篇

非标记定量法对乙醇诱导肝损伤的蛋白质组学研究

王小岳, 王道光, 郭继生, 李翠玲, 荆瑞瑞, 吕鑫, 陈新骏, 杨静华   

  1. 山东大学癌症研究中心, 山东 济南 250012
  • 收稿日期:2014-10-30 修回日期:2014-12-25 出版日期:2015-03-10 发布日期:2015-03-10
  • 通讯作者: 杨静华, E-mail:jyang@bu.edu E-mail:jyang@bu.edu

Label-free quantification of proteomic changes in ethanol-induced hepatocyte

WANG Xiaoyue, WANG Daoguang, GUO Jisheng, LI Cuiling, JING Ruirui, LÜ Xin, CHEN Xinjun, YANG Jinghua   

  1. Cancer Research Center, Shandong University, Jinan 250012, Shandong, China
  • Received:2014-10-30 Revised:2014-12-25 Online:2015-03-10 Published:2015-03-10

摘要: 目的 运用基于质谱的非标记定量法探讨乙醇处理的肝癌细胞蛋白质组学的变化。方法 将乙醇诱导损伤的肝癌细胞HepG2在溶液内酶解, 通过液质联用(LC-MS)检测酶解肽段, 应用非标定量软件Progenesis LC-MS分析蛋白表达的差异, 差异蛋白进一步应用蛋白分析数据库Panther 9.0和String 9.1统计分析。结果 Progenesis LC-MS分析显示, 有309个检测到的蛋白质含量存在显著差异, Panther统计表明, 这些差异蛋白包含各种催化酶类、细胞信号通路蛋白和结构蛋白等, 涉及凋亡、免疫应答等细胞内的重要的生理反应, String显示这些蛋白间存在密切的相互联系。结论 基于质谱的非标定量检测法可以为乙醇诱导的肝损伤机制研究提供依据与参考。

关键词: 非标记定量, 质谱, 蛋白质组学, 乙醇, 肝细胞损伤

Abstract: Objective To explore the proteomic changes in ethanol-induced hepatocyte cell line by a label-free quantification method. Methods Hepatocellular carcinoma cell line HepG2 was treated with ethanol and underwent in-solution-digestion. The total peptides were detected by liquid chromatography-mass spectrometry (LC-MS) and analyzed by Progenesis LC-MS, a commercial label-free quantification software. The differentially expressed proteins were further counted by Panther 9.0 and String 9.1 databases. Results More than 309 proteins were defined with significant difference in amount by Progenesis LC-MS. They were belong to catalytic enzymes, cell-signaling-related proteins, structural proteins, and soon which were involved in a variety of intracellular physiological responses, such as apoptosis and immune response. String showed there was a close relationship among these proteins. Conclusion Label-free quantification is an effective way of researching the mechanism of ethanol-induced hepatocyte damage.

Key words: Mass spectrometry, Label-free quantification, Proteomics, Hepatocyte damage, Ethanol

中图分类号: 

  • R394.3
[1] Zhang X, Fang AQ, Catherine PR, et al. Multi-dimensional liquid chromatography in proteomics-a review[J]. Anal Chim Acta, 2010, 664(2):101-113.
[2] Woods AG, Iosifescu DV, Darie CC. Biomarkers in major depressive disorder:the role of mass spectrometry[J]. Adv Exp Med Biol, 2014, 806:545-560.
[3] Zubiri I, Posada-Ayala M, Sanz-Maroto A, et al. Diabetic nephropathy induces changes in the proteome of human urinary exosomes as revealed by label-free comparative analysis[J]. J Proteomics, 2014, 16(96):92-102.
[4] de Graaf EL, Giansanti P, Altelaar AF, et al. Single-step enrichment by Ti4+-IMAC and label-free quantitation enables in-depth monitoring of phosphorylation dynamics with high reproducibility and temporal resolution[J]. Mol Cell Proteomics, 2014, 13(9):2426-2434.
[5] Willison KR, Klug DR. Quantitative single cell and single molecule proteomics for clinical studies[J]. Curr Opin Biotechnol, 2013, 24(4):745-751.
[6] Théron L, Gueugneau M, Coudy C, et al. Label-free quantitative protein profiling of vastus lateralis muscle during human aging[J]. Mol Cell Proteomics, 2014, 13(1):283-294.
[7] 陈晓超, 刘树滔, 陈躬瑞, 等. 酒精性肝炎的治疗进展[J]. 世界华人消化杂志, 2007, 15(10):1130-1136. CHEN Xiaochao, LIU Shutao, CHEN Gongrui, et al. Advances in treatment of alcoholic hepatitis[J]. World J Dig, 2007, 15(10):1130-1136.
[8] You Y, Yoo S, Yoon HG, et al. In vitro and in vivo hepatoprotective effects of the aqueous extract from Taraxacum officinale (dandelion) root against alcohol-induced oxidative stress[J]. Food Chem Toxicol, 2010, 48(6):1632-1637.
[9] Pochareddy S, Edenberg HJ. Chronic alcohol exposure alters gene expression in HepG2 cells[J]. Alcohol Clin Exp Res, 2012, 36(6):1021-1033.
[10] García-Ruiz C, Kaplowitz N, Fernandez-Checa JC. Role of mitochondria in alcoholic liver disease[J]. Curr Pathobiol Rep, 2013, 1(3):159-168.
[11] Chen NZ, Geng QQ, Zheng JB, et al. Suppression of the TGF-β/Smad signaling pathway and inhibition of hepatic stellate cell proliferation play a role in the hepatoprotective effects of curcumin against alcohol-induced hepatic fibrosis[J]. Int J Mol Med, 2014, 34(4):1110-1116.
[12] Li YY, Gao C, Shi YR, et al. Carbon monoxide alleviates ethanol-induced oxidative damage and inflammatory stress through activating p38 MAPK pathway[J]. Toxicol Appl Pharmacol, 2013, 273(1):53-58.
[13] Petrasek J, Iracheta-Vellve A, Csak T, et al. STING-IRF3 pathway links endoplasmic reticulum stress with hepatocyte apoptosis in early alcoholic liver disease[J]. Proc Natl Acad Sci U S A, 2013, 110(41):16544-16549.
[14] Zhong J, Kim MS, Chaerkady R, et al. TSLP signaling network revealed by SILAC-based phosphoproteomics[J]. Mol Cell Proteomics, 2012, 11(6):M112.017764. doi:10.1074/mcp.M112.017764
[15] Pütz SM, Boehm AM, Stiewe T, et al. iTRAQ analysis of a cell culture model for malignant transformation, including comparison with 2D-PAGE and SILAC[J]. J Proteome Res, 2012, 11(4):2140-2153.
[16] Wong JW, Cagney G. An overview of label-free quantitation methods in proteomics by mass spectrometry[J]. Methods Mol Biol, 2010, 604:273-283.
[17] Zhou W, Liotta LA, Petricoin EF. The spectra count label-free quantitation in cancer proteomics[J]. Cancer Genomics Proteomics, 2012, 9(3):135-142.
[18] Kannaste O, Suomi T, Salmi J, et al. Cross-correlation of spectral count ranking to validate quantitative proteome measurements[J]. J Proteome Res, 2014, 13(4):1957-1968.
[1] 刘超,刁婷婷,张红吉,刘红伟,杨永锐,李晓非. 飞行时间质谱在耐药结核病检测中的应用价值[J]. 山东大学学报 (医学版), 2024, 62(4): 61-67.
[2] 梁中昊,庄向华,黄珊,韩晓琳,华梦羽,琚丽萍,陈诗鸿. 饮食诱导肥胖雌鼠棕色脂肪组织蛋白质组学分析[J]. 山东大学学报 (医学版), 2024, 62(2): 10-19.
[3] 于丹凤,曹娟,于世佳,杜鲁涛,侯新国. 电化学发光法与液相色谱-串联质谱法评估糖尿病患者维生素D缺乏程度的效能比较与界值确定[J]. 山东大学学报 (医学版), 2023, 61(7): 90-95.
[4] 包舒晴,杨明月,刘端瑞,汪运山,郏雁飞. NOX4在幽门螺旋杆菌诱导胃癌细胞ROS中的作用[J]. 山东大学学报 (医学版), 2022, 60(6): 19-25.
[5] 李雁儒,李娟,李培龙,杜鲁涛,王传新. 胰腺癌不同进展期血清外泌体蛋白质组学分析[J]. 山东大学学报 (医学版), 2022, 60(10): 33-41.
[6] 关恒云,刘岚铮,姜慧钰,白爱英,刘辉,赵小冬,高尚,李娜,阮师漫. 利用时间质谱检出312例6月龄内健康婴儿鼻咽部微生态菌群的分布特征[J]. 山东大学学报 (医学版), 2022, 60(10): 49-56.
[7] 陈立晓,英信江,陈歆维,王菲,孙臻峰,董频. 下咽鳞癌蛋白质谱鉴定及预后靶分子筛选[J]. 山东大学学报 (医学版), 2021, 59(9): 140-147.
[8] 张华宇,殷思源,刘健,马嘉旭,宋茹,曹国起,王一兵. 氧糖剥夺条件下培养表皮干细胞的定量蛋白质组学分析[J]. 山东大学学报 (医学版), 2021, 59(4): 17-27.
[9] 刘冬科,王凤山. HILIC-MS/MS法鉴别肝素和硫酸乙酰肝素[J]. 山东大学学报 (医学版), 2021, 59(3): 41-47.
[10] 向琳,陈腊梅,王婧雯,李海铭,李浩宇,王菊,范玉琛,王凯. 30名健康志愿者饮用饮久舒对肝代谢酶的影响[J]. 山东大学学报 (医学版), 2021, 59(3): 81-85.
[11] 徐红照,郝宗山, 沈德嫱,李朝苹,陈立勇. 核糖体蛋白S6激酶2沉默在乙醇诱导的心肌胰岛素抵抗中的作用[J]. 山东大学学报 (医学版), 2020, 58(4): 84-89.
[12] 关新媛,高计芳,郭娟娟,刘师莲. 维生素D对大鼠肾上腺嗜铬细胞瘤PC12细胞中神经酰胺水平的影响[J]. 山东大学学报 (医学版), 2017, 55(12): 13-17.
[13] 刘亚飞, 张奇舒, 袁首道, 陈健行. HPV永生化细胞系与SiHa细胞系蛋白质表达差异的质谱分析[J]. 山东大学学报(医学版), 2015, 53(3): 22-26.
[14] 孙洪祥, 陈萍, 焦泽沼, 岳苏, 金天云, 向兰. 益智仁盐炙前后挥发油的GC-MS分析及抗乙酰胆碱酯酶活性测定[J]. 山东大学学报(医学版), 2015, 53(12): 27-32.
[15] 袁首道, 王凤琴, 张娇, 赵蔚明, 陈健行. 宫颈腺癌细胞与宫颈鳞癌细胞蛋白表达差异的质谱分析[J]. 山东大学学报(医学版), 2014, 52(8): 14-21.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!