基于血清药物化学及蛋白芯片技术探讨加味六君子汤治疗慢性萎缩性胃炎的作用机制  

Exploration of the Mechanism of Modified Liujunzi Decoction in Treating Chronic Atrophic Gastritis Based on Serum Pharmacochemistry and Protein Chip Technology

作　　者：张帅 周明[3] 宋沛祥 刘雪 邓雅依[1,2] 王颖 蔡皓 ZHANG Shuai;ZHOU Ming;SONG Peixiang;LIU Xue;DENG Yayi;WANG Ying;CAI Hao(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023 Jiangsu,China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing,Nanjing University of Chinese Medicine,Nanjing 210023 Jiangsu,China;School of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Nanjing 210023 Jiangsu,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南京中医药大学国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023 [3]南京中医药大学中医学院,江苏南京210023

出　　处：《中药新药与临床药理》2025年第3期357-367,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：江苏省自然科学基金项目(BK20210687)。

摘　　要：目的基于血清药物化学及蛋白芯片技术探讨加味六君子汤治疗慢性萎缩性胃炎的作用机制。方法(1)将Wistar大鼠随机分成空白对照组、加味六君子汤组(43.36 g·kg^(-1)),每日灌胃给药2次,持续7 d,制备加味六君子汤含药血清及空白血清。基于血清药物化学方法,采用超高效液相色谱-四极杆飞行时间质谱(UHPLC-Q-TOF-MS/MS)分析技术,并结合文献报道和数据库信息,确认加味六君子汤的入血成分。(2)使用PharmMapper网站进行入血成分作用靶点预测;利用GeneCards、OMIM、PharmGkb、TTD和DrugBank数据库筛选慢性萎缩性胃炎疾病相关靶点;将上述靶点输入韦恩图制作平台,所得交集靶点即为加味六君子汤治疗慢性萎缩性胃炎的潜在作用靶点。通过STRING数据平台对潜在作用靶点进行蛋白互作(PPI)网络构建及核心靶点筛选。通过Cytoscape软件导入加味六君子汤入血成分、潜在作用靶点,构建“入血成分-靶点”网络,筛选核心成分。通过DAVID数据库对潜在作用靶点进行GO功能及KEGG通路富集分析。利用AutoDock Vina软件对核心成分和核心靶点进行分子对接验证。(3)采用CCK-8法检测GES-1细胞活性,筛选1-甲基-3-硝基-1-亚硝基胍(MNNG)最合适的造模浓度及加味六君子汤含药血清的最佳给药浓度。采用40μmol·L^(-1)MNNG干预GES-1细胞,构建慢性萎缩性胃炎细胞模型,同时以20%含药血清干预24 h后,进行CSP100 plus磷酸化抗体芯片检测。结果(1)共鉴定出加味六君子汤入血成分23种。得到入血成分对应的作用靶点424个,慢性萎缩性胃炎疾病相关靶点1028个,取交集得到加味六君子汤治疗慢性萎缩性胃炎的潜在作用靶点109个。筛选得到加味六君子汤治疗慢性萎缩性胃炎的核心靶点:TP53、IL6、TNF、SRC、EGFR、AKT1、CTNNB1、MMP9、CASP3、MAPK8,以及核心成分:甘草酸、DL-精氨酸、芸香柚皮苷、大豆素、茯苓酸G、人参皂苷Rg2、人参皂�Objective To explore the mechanism of Modified Liujunzi Decoction in the treatment of chronic atrophic gastritis based on serum pharmacochemistry and protein chip technology.Methods(1)Wistar rats were randomly divided into blank control group and Modified Liujunzi Decoction group(43.36 g·kg^(-1)),intragastric administration was given two times a day for consecutive 7 days to prepare Modified Liujunzi Decoction containing serum and blank serum.Based on the serum pharmacochemistry method,the ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry(UHPLC-Q-TOF-MS/MS)technology combined with literature reports and database information was used to confirm the constituents absorbed into blood of Modified Liujunzi Decoction.(2)The targets of the constituents absorbed into blood were predicted using PharmMapper website.Disease targets related to chronic atrophic gastritis were screened using GeneCards,OMIM,PharmGkb,TDD,and DrugBank databases.The above targets were input into the Venn diagram production platform,and the obtained intersection targets were the potential targets of Modified Liujunzi Decoction in the treatment of chronic atrophic gastritis.Protein-protein interaction(PPI)network construction and core target screening of potential targets were performed by STRING data platform.The constituents absorbed into blood and potential targets of Modified Liujunzi Decoction were introduced by Cytoscape software,and the‘constituents absorbed into blood-targets’network was constructed to screen the core components.GO function and KEGG signaling pathway enrichment analysis of potential targets were performed by DAVID database.AutoDock Vina software was used to verify the molecular docking of core components and core targets.(3)The CCK-8 method was used to detect the activity of GES-1 cells,and the most suitable modeling concentration of 1-methyl-3-nitro-1-nitrosoguanidine(MNNG)and the optimal administration concentration of Modified Liujunzi Decoction containing serum were screened.GES

关 键 词：加味六君子汤 慢性萎缩性胃炎 血清药物化学 网络药理学 蛋白芯片技术 PI3K/AKT信号通路 MAPK信号通路 GES-1人胃黏膜上皮细胞 大鼠血清 

分 类 号：R285.5[医药卫生—中药学]