应用iTRAQ技术筛选右归丸对肾阳虚证干预效应的血清差异表达蛋白  

Screening for differentially expressed serum proteins of Kidney-yang deficiency after Regulated by Yougui Pills using iTRAQ technology

作　　者：谭从娥[1] 黄祥云[1] 郭小青[1] 

机构地区：[1]陕西中医学院,陕西咸阳712046

出　　处：《时珍国医国药》2014年第9期2277-2279,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81072731)

摘　　要：目的采用核素标记相对和绝对定量(isobaric Tags for Relative and Absolute Quantitation,iTRAQ)蛋白组学技术分析肾阳虚证患者右归丸治疗前后比较的血清差异表达蛋白。方法 4例肾阳虚证患者给予右归丸口服一个月,于治疗前后分别采取外周血,取血清提取蛋白,进行蛋白变性还原及酶解,最后进行iTRAQ标记和Q-Exactive质谱仪鉴定,质谱分析原始数据用软件Mascot 2.2和Proteome Discoverer 1.4进行查库鉴定及定量分析,筛选并鉴定差异表达蛋白,并对差异表达蛋白进行生物信息学分析。结果共鉴定血清蛋白236个,有定量信息的171个,其中在4组中均差异表达且表达方向一致的蛋白有6个,上调4个,下调2个。6种差异表达蛋白参与22种生物学过程和1条京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路。结论补体C3(Complement C3,C3)、半胱氨酸抑制酶C(Cystatin-C,Cys-C)、载脂蛋白C-Ⅲ(apolipoprotein C-Ⅲ,APOC3)、载脂蛋白E(apolipoprotein E,APOE)等蛋白可能与右归丸的疗效的发挥密切相关。Objective To screen differentially expressed serum proteins of the Kidney-yang deficiency associated with Yougui Pills. Methods Four patients with kidney-yang deficiency were selected for 1 month of Yougui Pills taken treatment. Accumulated clinical scores of Kidney-yang deficiency were assessed pre-treatment and post-treatment using a 40-factor questionnaire of Kidney-yang deficiency. 5ml Blood was collected before and after Yougui Pills treatment and protein was extracted from serum. After albuminous degeneration,reduction and enzymolysis were performed,the iTRAQ was adopted for labeling and Q-Exactive mass spectrometer was for identification. The original data were searched with MASCOTT 2. 2 and Proteome Discoverer 1. 4 to achieve the expression difference. Results A total of 236 serum proteins were identified,of them,171 proteins reached the strict quantitative standard. Six serum proteins were found differentially expressed after Yougui Pills regulated,4 were up-regulated and2 down-regulated. These differentially expressed proteins are enriched in 22 biological processes and 1 KEGG pathway. Conclusion Serum proteins of C3,Cys-C,APOC3,and APOE may be the biological marker of Yougui Pills for patients with Kidney-yang deficiency.

关 键 词：肾阳虚证 右归丸 ITRAQ 蛋白质组 

分 类 号：R2-03[医药卫生—中医学]