基于蛋白质组学探讨仙连解毒方干预小鼠结直肠癌湿热瘀毒证模型的作用靶点及信号通路研究  

Study on the Target and Signaling Pathway of Xian-Lian-Jie-Du Decoction in Mouse Model of Colorectal Cancer with Damp-heat and Stagnant Toxin Syndrome Based on Proteomics

作　　者：陆思丞 程海波[1,2] 余成涛 沈卫星 徐长亮[1,2] 谭佳妮 赖岳阳[1,2] 范旻旻 Lu Sicheng;Cheng Haibo;Yu Chengtao;Shen Weixing;Xu Changliang;Tan Jiani;Lai Yueyang;Fan Minmin(The First Clinical Medical College,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor,Nanjing 210023,China)

机构地区：[1]南京中医药大学第一临床医学院,南京210023 [2]江苏省中医药防治肿瘤协同创新中心,南京210023

出　　处：《世界科学技术-中医药现代化》2022年第8期3067-3076,共10页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：国家自然科学基金委员会重点项目(81930117):基于肿瘤微环境的晚期大肠癌湿热瘀毒证及消癌解毒方抑制转移的生物学基础研究,负责人:程海波

摘　　要：目的应用蛋白质组学方法探讨仙连解毒方(Xian-Lian-Jie-Du Decotion,XLJDD)治疗小鼠结直肠癌湿热瘀毒证模型的作用靶点及信号通路。方法采用偶氮甲烷(AOM)/葡聚糖硫酸钠(DSS)结合高糖高脂饲料喂养的复合方法建立小鼠结直肠癌湿热瘀毒证模型,将小鼠分为空白对照(Control)组、湿热瘀毒证模型(Model)组、仙连解毒方处理(XLJDD)组。XLJDD组于造模同时开始给药(12.9 g·kg^(-1)),每周6次,共112天,末次给药4 h后,收集各组小鼠结直肠组织,迅速液氮保存。蛋白质质谱检测采用非数据依赖型采集模式(DIA),通过Protein Discovery 2.2软件进行蛋白质搜库鉴定,对蛋白及肽段信息定量分析,筛选各组差异表达蛋白,并对差异蛋白进行GO(Gene Oncology)和KEGG(Kyoto Encyclopedia of Genes and Genomes)富集分析。结果Model组与Control组相比,共有差异表达蛋白267个,XLJDD组与Model组相比,共有225个差异表达蛋白,GO以及KEGG富集分析发现仙连解毒方干预后,发现这些蛋白主要定位在细胞膜上,能够发挥细胞调节功能等分子功能,参与细胞粘附、Th1/Th2细胞分化、Th17细胞分化和细胞衰老通路等生物学进程相关。结论通过蛋白质组学研究提示,XLJDD可能通过调节免疫细胞功能,发挥干预小鼠结直肠癌湿热瘀毒证的作用。Objective To explore the target and signaling pathway of Xian-Lian-Jie-Du Decoction(XLJDD)in mouse model of colorectal cancer with damp-heat and stagnant toxin syndrome by proteomics.Methods The mice model was established by the compound method of azomethane(AOM)/sodium glucan sulfate(DSS)fed with high fat and high glucose diet at the same time.These mice were divided into control group(Control),damp-heat and stagnant toxin syndrome model group(Model)and Xian-Lian-Jie-Du Decoction group(XLJDD).The XLJDD group mice were given intragastric administration(12.9 g·kg^(-1))at the beginning of modeling,six times a week for 112 days.The colorectal tissues of each group were collected and stored in liquid nitrogen after 4 h last the administration.The mass spectrometry was performed using data-independent acquisition(DIA),then protein and peptide were identified by Protein Discovery2.2 software and quantified to screen differentially expressed proteins in each group.The differential proteins were analyzed by GO(Gene Oncology)and KEGG(Kyoto Encyclopedia of Genes and Genomes).Results There were 267differentially expressed proteins in Model compared with Control.Later,there were 225 differentially expressed proteins after intervention with XLJDD.In addition,GO and KEGG enrichment analysis showed that differentially expressed proteins differential proteins after intervention with XLJDD were mainly located on the cell membrane and played a role in molecular functions such as in Th1/Th2 cell differentiation,Th17 cell differentiation and cellular senescence pathway.Conclusion Proteomic studies suggest that XLJDD may play a role in the intervention of damp-heat stasis toxin syndrome of colorectal cancer in mice by regulating the function of immune cells.

关 键 词：结直肠癌 仙连解毒方 湿热瘀毒证 蛋白质组学 

分 类 号：R285.5[医药卫生—中药学]