Abstract：
BACKGROUND: Functional dyspepsia (FD) is a prevalent gastrointestinal disorder characterised by high incidence and recurrence rates, posing significant health risks. Erpixing Granules (EPX), approved by the National Food and Drug Administration in 2002, are known for their spleen and stomach invigorating properties, effectively treating FD. However, its mechanism of action remains unclear.
OBJECTIVE: This study aims to elucidate EPX\'s mechanism of treating FD through network pharmacology, and experimental validation using FD animal models.
METHODS: In this study, the chemical composition of EPX in positive and negative ion modes was analyzed by UHPLC-Q-TOF MS. The mass spectral data were processed and analyzed using MS-DIAL software to automatically match compound fragment information and identify the known components with the compound database to obtain the active components of EPX. SwissTargetPrediction was used to obtain EPX targets, while FD-related targets were sourced from GeneCards, OMIM and DisGeNET databases. A protein-protein interaction (PPI) network was constructed using the STRING platform, and potential signalling pathways of EPX were determined through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Finally, an FD model was established in rates by administering a 0.1% iodoacetamide sucrose solution, followed by tail clamp stimulation to experimentally validate the network pharmacology findings.
RESULTS: Our results revealed 139 effective ingredients in EPX, targeting 60 core FD-related genes. PPI network analysis identified EGFR, CTNNB1 and NFκB1 as core target genes. The KEGG pathway analysis indicated that EPX can modulate FD progression through the PI3K/AKT signalling pathway. Animal experiments demonstrated EPX\'s capacity to increase body mass, food intake and food utilisation efficiency in FD rats, alongside increased gastric juice secretion, pepsin activity, trypsin activity, cholesterol, bile acid and bilirubin activity. HE examination revealed that EPX improved the inflammatory infiltration of gastric mucosal cells in rats. Furthermore, EPX also promoted gastric emptying and intestinal propulsion in mice. These results suggest that EPX improves spleen and stomach function, enhances the protective effect on the spleen and stomach and promotes food digestion and absorption. Immunofluorescence studies revealed upregulated expression of PI3K, AKT and ANO1 proteins in gastric tissue following EPX administration, while Western blotting indicated increased expression of SCF and C-kit proteins.
CONCLUSIONS: Suggesting EPX\'s anti-FD effect may involve the regulation of the SCF/C-kit signalling pathway and activation of downstream PI3K/AKT signalling pathway, thereby promoting gastrointestinal motility and improving FD symptoms.
OBJECTIVE: This study aims to elucidate EPX\'s mechanism of treating FD through network pharmacology, and experimental validation using FD animal models.
METHODS: In this study, the chemical composition of EPX in positive and negative ion modes was analyzed by UHPLC-Q-TOF MS. The mass spectral data were processed and analyzed using MS-DIAL software to automatically match compound fragment information and identify the known components with the compound database to obtain the active components of EPX. SwissTargetPrediction was used to obtain EPX targets, while FD-related targets were sourced from GeneCards, OMIM and DisGeNET databases. A protein-protein interaction (PPI) network was constructed using the STRING platform, and potential signalling pathways of EPX were determined through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Finally, an FD model was established in rates by administering a 0.1% iodoacetamide sucrose solution, followed by tail clamp stimulation to experimentally validate the network pharmacology findings.
RESULTS: Our results revealed 139 effective ingredients in EPX, targeting 60 core FD-related genes. PPI network analysis identified EGFR, CTNNB1 and NFκB1 as core target genes. The KEGG pathway analysis indicated that EPX can modulate FD progression through the PI3K/AKT signalling pathway. Animal experiments demonstrated EPX\'s capacity to increase body mass, food intake and food utilisation efficiency in FD rats, alongside increased gastric juice secretion, pepsin activity, trypsin activity, cholesterol, bile acid and bilirubin activity. HE examination revealed that EPX improved the inflammatory infiltration of gastric mucosal cells in rats. Furthermore, EPX also promoted gastric emptying and intestinal propulsion in mice. These results suggest that EPX improves spleen and stomach function, enhances the protective effect on the spleen and stomach and promotes food digestion and absorption. Immunofluorescence studies revealed upregulated expression of PI3K, AKT and ANO1 proteins in gastric tissue following EPX administration, while Western blotting indicated increased expression of SCF and C-kit proteins.
CONCLUSIONS: Suggesting EPX\'s anti-FD effect may involve the regulation of the SCF/C-kit signalling pathway and activation of downstream PI3K/AKT signalling pathway, thereby promoting gastrointestinal motility and improving FD symptoms.
摘要：
背景：功能性消化不良（FD）是一种常见的胃肠道疾病，其特点是发病率和复发率高，构成重大健康风险。Erpixing颗粒（EPX），2002年由国家食品药品监督管理局批准，以其健脾和胃的特性而闻名，有效治疗FD。然而,其作用机制尚不清楚。
目的：本研究旨在阐明EPX通过网络药理学治疗FD的机制，并使用FD动物模型进行实验验证。
方法：在本研究中，通过UHPLC-Q-TOFMS分析了正负离子模式下EPX的化学成分。使用MS-DIAL软件对质谱数据进行处理和分析,以自动匹配化合物片段信息并将已知组分与化合物数据库进行识别,从而获得EPX的活性组分。SwissTargetPrediction用于获得EPX目标，虽然FD相关目标来自GeneCards，OMIM和DisGeNET数据库。使用STRING平台构建了蛋白质-蛋白质相互作用(PPI)网络，通过基因本体论（GO）和京都基因和基因组百科全书（KEGG）富集分析确定了EPX的潜在信号通路。最后,通过给予0.1%碘乙酰胺蔗糖溶液建立FD模型，然后进行尾部钳夹刺激，以实验验证网络药理学发现。
结果：我们的结果显示EPX中有139种有效成分，靶向60个核心FD相关基因。PPI网络分析确定了EGFR，CTNNB1和NFκB1为核心靶基因。KEGG通路分析表明EPX可以通过PI3K/AKT信号通路调节FD进展。动物实验证明了EPX增加体重的能力，FD大鼠的食物摄入量和食物利用效率，随着胃液分泌增加，胃蛋白酶活性，胰蛋白酶活性，胆固醇,胆汁酸和胆红素活性。HE检查显示EPX改善了大鼠胃粘膜细胞的炎性浸润。此外，EPX还促进小鼠胃排空和肠推进。这些结果表明，EPX可以改善脾胃功能，增强对脾胃的保护作用，促进食物消化吸收。免疫荧光研究显示PI3K的表达上调，EPX给药后胃组织中的AKT和ANO1蛋白，而Western印迹显示SCF和C-kit蛋白的表达增加。
结论：提示EPX的抗FD作用可能涉及SCF/C-kit信号通路的调节和下游PI3K/AKT信号通路的激活，从而促进胃肠蠕动和改善FD症状。
目的：本研究旨在阐明EPX通过网络药理学治疗FD的机制，并使用FD动物模型进行实验验证。
方法：在本研究中，通过UHPLC-Q-TOFMS分析了正负离子模式下EPX的化学成分。使用MS-DIAL软件对质谱数据进行处理和分析,以自动匹配化合物片段信息并将已知组分与化合物数据库进行识别,从而获得EPX的活性组分。SwissTargetPrediction用于获得EPX目标，虽然FD相关目标来自GeneCards，OMIM和DisGeNET数据库。使用STRING平台构建了蛋白质-蛋白质相互作用(PPI)网络，通过基因本体论（GO）和京都基因和基因组百科全书（KEGG）富集分析确定了EPX的潜在信号通路。最后,通过给予0.1%碘乙酰胺蔗糖溶液建立FD模型，然后进行尾部钳夹刺激，以实验验证网络药理学发现。
结果：我们的结果显示EPX中有139种有效成分，靶向60个核心FD相关基因。PPI网络分析确定了EGFR，CTNNB1和NFκB1为核心靶基因。KEGG通路分析表明EPX可以通过PI3K/AKT信号通路调节FD进展。动物实验证明了EPX增加体重的能力，FD大鼠的食物摄入量和食物利用效率，随着胃液分泌增加，胃蛋白酶活性，胰蛋白酶活性，胆固醇,胆汁酸和胆红素活性。HE检查显示EPX改善了大鼠胃粘膜细胞的炎性浸润。此外，EPX还促进小鼠胃排空和肠推进。这些结果表明，EPX可以改善脾胃功能，增强对脾胃的保护作用，促进食物消化吸收。免疫荧光研究显示PI3K的表达上调，EPX给药后胃组织中的AKT和ANO1蛋白，而Western印迹显示SCF和C-kit蛋白的表达增加。
结论：提示EPX的抗FD作用可能涉及SCF/C-kit信号通路的调节和下游PI3K/AKT信号通路的激活，从而促进胃肠蠕动和改善FD症状。
