| 引用本文: | 李彦,朱敏,曹雯,胡仁统,张学荣,廖明.基于iTRAQ技术分析联合用药作用大鼠肝星状细胞后差异表达的蛋白[J].中国现代应用药学,2014,31(11):1302-1307. |
| LI Yan,ZHU Min,CAO Wen,HU Rentong,ZHANG Xuerong,LIAO Ming.Analysis of the Differentially Expressed Proteins on Combination-therapy in Rat Hepatic Stellate Cells by iTRAQ[J].Chin J Mod Appl Pharm(中国现代应用药学),2014,31(11):1302-1307. |
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| 摘要: |
| 目的 筛选联合用药作用大鼠肝星状细胞(HSC-T6)后差异表达的蛋白质，以探讨联合用药抑制肝星状细胞增殖的机制。方法 以牛磺酸、表没食子儿茶素没食子酸酯(EGCG)和三羟基异黄酮三者组成的联合用药作用HSC-T6后，提取细胞总蛋白，运用同位素标记的iTRAQ技术结合高效液相色谱-电喷雾串联质谱(LC-ESI-MS/MS)的定量蛋白质组学方法，分析和鉴定差异表达的蛋白质，运用生物信息学分析差异表达的蛋白质功能。结果 质谱共分析鉴定到727种蛋白质。3次实验分别鉴定到的差异表达蛋白质数量分别为85，102，94个，在3次重复均鉴定到的差异表达蛋白为52个，其中上调蛋白24个，下调蛋白28个。生物信息学分析表明，差异表达的蛋白参与翻译后修饰、转录、重组和信号传导等过程，在这些蛋白质中既有与肝纤维化发生、发展直接相关的蛋白质如基质金属蛋白酶抑制剂1、层黏连蛋白、前纤维蛋白2等，也有在蛋白质的相互作用中处于功能网络交叉点的蛋白质如细胞周期素依赖性激酶4、肝癌衍生生长因子、谷氨酸脱氢酶 1和丝氨酸/精氨酸剪接因子9等。结论 联合用药抑制肝星状细胞增殖的作用，可能是通过调控多种蛋白质的表达来实现，所获得的差异表达蛋白可能在肝纤维化发生发展和药物抗肝纤维化作用过程中扮演重要角色。 |
| 关键词: 联合用药 肝星状细胞 细胞蛋白 iTRAQ |
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| 基金项目:国家自然科学基金项目(81160063)；广西自然科学基金项目(2013GXNSFBA019167)；广西医学科学实验中心开放基金专项项目(KFJJ2010-41) |
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| Analysis of the Differentially Expressed Proteins on Combination-therapy in Rat Hepatic Stellate Cells by iTRAQ |
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LI Yan1, ZHU Min1, CAO Wen2, HU Rentong2, ZHANG Xuerong2, LIAO Ming2
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1.Guangxi University Library, College of Animal Science and Technology of Guangxi University, Nanning 530004, China;2.Key Laboratory Joint Established by Ministry of Education and Guangxi, Key Laboratory of Guangxi, Medical Scientific Research Center of Guangxi Medical University, Nanning 530021, China
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| Abstract: |
| OBJECTIVE To detect protein expression differences after combination-therapy on HSC-T6 and to explore antifibrotic mechanism of combination-therapy based on the protein level. METHODS HSC-T6 as target cells, cells proteins were extracted after treated by combination drug of taurine, epigallocatechin gallate (EGCG) and genistein. After equal amount of protein digested by trypsin, the iTRAQ labeled protein peptide, separated by the cation column, LC-ESI-MS/MS used to detect and identify differentially expressed proteins. The functions of proteins were analyzed by bioinformatics. RESULTS Total 727 proteins were identified by MS. Three biological replicates identified 85, 102 and 94 differentially expressed proteins respectively. The analysis manifest 52 differentially expressed proteins were identified in three biological replicates simultaneously, of which 24 differentially expressed proteins were up-regulated, 28 differentially expressed proteins were down-regulated. The differentially expressed proteins correlate with post-translational modification, transcription, recombination and signal transduction pathways. CONCLUSION Combined drug treatment with taurine, EGCG and genistein can impact HSC protein expression. The antifibrotic effects of this drug combination may be due to regulation of protein expression. |
| Key words: combination-therapy liver fibrosis cell proteins iTRAQ |
