基于iTRAQ技术的安胃汤干预胃溃疡肝郁脾虚证大鼠蛋白质组学研究  

iTRAQ Based Proteomics Study of Anwei Decoction on Gastric Ulcer with Liver Depression and Spleen Asthenia Syndrome in Rats

作　　者：罗蔚[1] 唐友明[1] 韩叶芬[1] 吴德坤[1] 牛豫洁[1] 夏泽[1] 韦维[1] 郑景辉[1] LUO Wei;TANG Youming;HAN Yefen;WU Dekun;NIU Yujie;XIA Ze;WEI Wei;ZHENG Jinghui(Ruikang Affiliated Hospital,Guangxi University of Chinese Medicine,Nanning 530011 Guangxi,China)

机构地区：[1]广西中医药大学附属瑞康医院,广西南宁530011

出　　处：《中药新药与临床药理》2019年第1期72-81,共10页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81360533);广西自然科学基金项目(2014GXNSFAA118140)

摘　　要：目的应用iTRAQ蛋白组学技术鉴定筛选安胃汤干预胃溃疡肝郁脾虚证大鼠胃黏膜细胞的差异表达蛋白,初步探讨安胃汤干预胃溃疡肝郁脾虚证大鼠的分子作用机制。方法采用乙酸法配合多因素复合模拟中医病因制备胃溃疡肝郁脾虚证大鼠模型,提取胃黏膜原代细胞进行培养,制备安胃汤含药血清对其进行干预,采用iTRAQ技术鉴定筛选差异蛋白。通过DAVID、KEGG、VISANT和STRING数据库对差异蛋白进行注释、富集分析、信号转导通路分析和蛋白质相互作用分析,并对其中的关键蛋白通过Western Blot实验进行验证。结果安胃汤组与对照组之间共有59个差异表达蛋白,生物信息学分析显示其主要参与53种生物学过程,10种细胞组分,16类分子途径,以及74条信号转导通路;Western Blot验证实验结果与iTRAQ蛋白组学结果基本一致。结论基于iTRAQ蛋白组学可筛选出安胃汤干预胃溃疡肝郁脾虚证大鼠胃黏膜细胞相关蛋白(Akt1、Jak1、Jak2、Plcb3、Adrb2、Stat2、Prkcg、Plcb4、Ptpn2、Chrm3及Ifngr1),初步分析提示其与胃溃疡的多种细胞生物学活动有关。Objective To detect protein expression differences in gastric mucosal cells with gastric ulcer with liverdepression and spleen-deficiency syndrome after treatment with Anwei decoction and to preliminarily explore the molecular biological mechanism. Methods The rat model of gastric ulcer with liver stagnation and spleen deficiency syndrome was prepared by acetic acid combined with multi-factor simulation method of TCM etiology. The gastric mucosal cells of the rat model were isolated and intervened by serum containing Anwei decoction. The altered proteins in gastric mucosal cells were identified by iTRAQ,the biochemical pathways and regulatory networks were analyzed with DAVID,GO,KEGG,VISANT and STRING databases. Key proteins were verified by Western Blot.Results Fifty-nine differentially expressed proteins were found between the Anwei decoction group and the control group. Bioinformatics analysis showed that 59 differentially expressed proteins were mainly involved in 53 biological processes, 10 cell components, 16 molecular pathways and 74 signal transduction pathways. Most results of Western Blot verification tests are the same as those of iTRAQ proteomics. Conclusion The differentially expressed proteins(Akt1,Jak1,Jak2,Plcb3,Adrb2,Stat2,Prkcg,Plcb4,Ptpn2,Chrm3 and Ifngr1) in gastric mucosal cells with gastric ulcer coupled liver-depression and spleen-deficiency syndrome intervened by Anwei decoction was screened by iTRAQ proteomics. The preliminary analysis suggests that it is related to various cellular biological activities of gastric ulcers,which is a mechanism worthy of further study for the action of Anwei decoction.

关 键 词：安胃汤 胃溃疡 胃黏膜细胞 肝郁脾虚证 蛋白质组学 生物信息学 

分 类 号：R285.5[医药卫生—中药学] R857.3[医药卫生—中医学]