基于质谱分析和网络药理学探究六君安胃方对化疗相关性腹泻小鼠小肠组织NF-κB与iNOS蛋白的影响  

Effects of Liujun Anwei Prescription on NF-κB and iNOS protein in small intestine of mice with chemotherapy-related diarrhea based on mass spectrometry and network pharmacology

作　　者：阴毅然 何斌[1] 赵娜 吴宏伟[2] 杨宇飞[1] Yin Yiran;He Bin;Zhao Na;Wu Hongwei;Yang Yufei(Oncology Department,Xiyuan Hospital,China Academy of Chinese Medical Science,Beijing 100091,China;Pharmacological Research Center,Institute of Traditional Chinese Medicine,China Academy of Chinese Medical Science,Beijing 100700,China)

机构地区：[1]中国中医科学院西苑医院肿瘤科,北京100091 [2]中国中医科学院中药研究所药理研究中心,北京100700

出　　处：《国际中医中药杂志》2023年第10期1259-1270,共12页International Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81973676);中国中医科学院科技创新工程(CI2021A01812,CI2021B009);国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202205)。

摘　　要：目的探究六君安胃方防治化疗相关性腹泻的潜在关键靶点及其对化疗相关性腹泻小鼠小肠组织NF-κB与iNOS蛋白的影响,揭示其抗炎止泻活性成分及分子作用机制。方法采用高分辨质谱技术对六君安胃方的化学成分进行定性分析。检索PubChem数据库获得六君安胃方活性成分,利用Swiss Target Prediction数据库进行靶点预测;检索DisGeNET、OMIM、GeneCards数据库获得化疗相关性腹泻相关靶点;将六君安胃方活性成分靶点与腹泻疾病相关的靶点取交集,得到六君安胃方治疗化疗相关性腹泻的潜在靶点;采用Cytoscape 3.7.1软件构建PPI网络及“成分-靶点-通路”网络;DAVID数据库对交集靶点进行GO功能、KEGG通路富集分析;利用Autodock软件将网络中预测到的潜在活性成分和靶点进行分子对接验证。将60只C57BL/6J雄性小鼠按随机数字表法分为空白组、模型组、阳性对照组及六君安胃方高、中、低剂量组,每组10只。除空白组外,其余各组小鼠腹腔注射氟尿嘧啶注射液50 mg/kg制备化疗相关性腹泻小鼠模型。各组给予相应药物干预14 d后,采用Western blot法检测小鼠空肠NF-κB、iNOS蛋白表达。结果共鉴定出197个化学成分,经筛选得到156个六君安胃方防治化疗相关腹泻的关键成分,涉及82个潜在作用靶点,主要通过NOS2等关键靶点,癌症通路、PI3K-Akt、NF-κB信号通路发挥作用。实验结果显示,六君安胃方可降低化疗相关性腹泻小鼠肠组织NF-κB、iNOS蛋白表达。结论揭示了六君安胃方的药效物质基础,可通过降低NF-κB及iNOS水平,影响肠道组织的炎症反应,改善肠黏膜屏障功能,进而改善化疗相关性腹泻。Objective To investigate the potential key targets of Liujun Anwei Prescription and its effects on NF-κB/iNOS-NO in small intestine of mice with chemotherapy-associated diarrhea;To reveal the anti-inflammatory components and molecular mechanism.Methods UPLC-Q/TOF MS combined with UNIFI software was used to analyze the chemical components of Liujun Anwei Prescription.PubChem database was searched to obtain the active components of Liujun Anwei Prescription,and the Swiss Target Prediction was used to predict the targets.The database of DisGeNET,OMIM and GeneCards were searched to obtain the targets of chemotherapy-related diarrhea.The potential targets of Liujun Anwei Prescription in the treatment of chemotherapy-related diarrhea diseases were obtained by crossing the targets of active components of Liujun Anwei Prescription and those related to diarrhea diseases.The PPI network and component-target-pathway network were constructed by Cytoscape 3.7.1 software,and the intersecting targets were analyzed by GO and KEGG based on David Database.The potential active components and potential targets predicted in the network were verified by using Autodock software.60 C57BL/6J male mice were divided into normal control group,model group,positive control group and Liujun Anwei Prescription high-,medium-and low-dosage groups according to random number table method,with 10 mice in each group.In addition to the normal control group,the other groups of mice were intraperitoneally injected with 5-fluorouracil injection 50 mg/kg preparation to construct CID mouse model.After 14 days,the expressions of NF-κB and iNOS in jejunum were detected by Western blot.Results A total of 197 compounds were identified,and 156 key compounds of Liujun Anwei Prescription were screened,involving 82 potential targets,mainly through NOS2 and other key targets,playing a role through cancer pathway,PI3K-Akt,NF-κB signal pathway.The experimental results showed that Liujun Anwei Prescription could significantly down-regulate the protein expressions o

关 键 词：质谱法 六君安胃方 化疗相关性腹泻 网络药理学 分子对接模拟 

分 类 号：R285.5[医药卫生—中药学]