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 40 µmol·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 1 028 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、人参皂苷 Ro、香风草甙、芹糖异甘草苷、人参皂苷 Rb1。潜在作用靶点涉及的生物过程主要有对氧化应 激的反应、脂多糖反应、细胞来源分子反应、凋亡信号通路等；KEGG 通路主要涉及病毒、癌症、炎症等方 面，主要通路包括 PI3K/AKT 信号通路、TNF 信号通路、IL-17 信号通路、MAPK 信号通路等。核心成分与核 心靶点的 100 组分子对接结果中，结合能≤ -5 kcal·mol-1的对接组合有 93 组，结合能≤ -7 kcal·mol-1的对接组 合有 73 组。（2）与模型组比较，经含药血清干预后，共有 49 个磷酸化抗体和 46 个非磷酸化抗体发生了明显变 化。其中，在 PI3K/AKT 信号通路中有 18 个磷酸化抗体和 11 个非磷酸化抗体发生了显著变化；在 MAPK 信号 通路中有 13 个磷酸化抗体和 17 个非磷酸化抗体发生了显著变化。结论 加味六君子汤可能通过甘草酸、人参 皂苷 Rg2、大豆素等入血成分，调控 PI3K/AKT、MAPK 关键信号通路，发挥治疗慢性萎缩性胃炎并延缓其向 胃癌转变的作用。

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-1 cells were treated with 40 µmol·L-1 MNNG to construct a cell model of chronic atrophic gastritis. At the same time， after 24 hours of intervention with 20% drug-containing serum， CSP100 plus phosphorylated antibody chip detection was performed. Results （1） A total of 23 constituents absorbed into blood of Modified Liujunzi Decoction were identified. A total of 424 targets corresponding to constituents absorbed into blood and 1 028 targets related to chronic atrophic gastritis were obtained，and 109 potential targets of Modified Liujunzi Decoction in the treatment of chronic atrophic gastritis were obtained. The core targets of Modified Liujunzi Decoction in the treatment of chronic atrophic gastritis were screened：TP53，IL6，TNF，SRC，EGFR，AKT1，CTNNB1，MMP9， CASP3，MAPK8，and core components：glycyrrhizic acid，DL-arginine，narirutin，daidzein，pachymic acid G， ginsenoside Rg2， ginsenoside Ro， vanillin， apioside， ginsenoside Rb1. The biological processes involved in the potential targets mainly include the response to oxidative stress，lipopolysaccharide response，cell-derived molecular response，apoptosis signaling pathway，etc. The KEGG pathway mainly involves viruses，cancer，inflammation，etc. The main pathways include PI3K/AKT signaling pathway，TNF signaling pathway，IL-17 signaling pathway，MAPK signaling pathway， etc. Among the 100 groups of molecular docking results of core components and core targets， there were 93 groups of docking combinations with binding energy ≤ -5 kcal·mol-1 and 73 groups of docking combinations with binding energy ≤ -7 kcal·mol-1 . （2） Compared with the model group，a total of 49 phosphorylated antibodies and 46 non-phosphorylated antibodies changed significantly after medicated serum intervention. Among them，18 phosphorylated antibodies and 11 non-phosphorylated antibodies were significantly changed in the PI3K/AKT signaling pathway； in the MAPK signaling pathway， 13 phosphorylated antibodies and 17 non-phosphorylated antibodies were significantly changed. Conclusion Modified Liujunzi Decoction may regulate the key signaling pathways of PI3K/AKT and MAPK through constituents absorbed into blood such as glycyrrhizic acid， ginsenoside Rg2 and daidzein，and play a role in treating chronic atrophic gastritis and delaying its transformation to gastric cancer.

R285.5

江苏省自然科学基金项目（BK20210687）