DataSheet_3_Material basis and molecular mechanisms of Chaihuang Qingyi Huoxue Granule in the treatment of acute pancreatitis based on network pharmacology and molecular docking-based strategy.zip

ObjectivesThis study aimed to analyze active compounds and signaling pathways of CH applying network pharmacology methods, and to additionally verify the molecular mechanism of CH in treating AP.
Materials and methodsNetwork pharmacology and molecular docking were firstly used to identify the active components of CH and its potential targets in the treatment of AP. The pancreaticobiliary duct was retrogradely injected with sodium taurocholate (3.5%) to create an acute pancreatitis (AP) model in rats. Histological examination, enzyme-linked immunosorbent assay, Western blot and TUNEL staining were used to determine the pathway and mechanism of action of CH in AP.
ResultsNetwork pharmacological analysis identified 168 active compounds and 276 target proteins. In addition, there were 2060 targets associated with AP, and CH had 177 targets in common with AP. These shared targets, including STAT3, IL6, MYC, CDKN1A, AKT1, MAPK1, MAPK3, MAPK14, HSP90AA1, HIF1A, ESR1, TP53, FOS, and RELA, were recognized as core targets. Furthermore, we filtered out 5252 entries from the Gene Ontology(GO) and 186 signaling pathways from the Kyoto Encyclopedia of Genes and Genomes(KEGG). Enrichment and network analyses of protein-protein interactions predicted that CH significantly affected the PI3K/AKT signaling pathway, which played a critical role in programmed cell death. The core components and key targets showed strong binding activity based on molecular docking results. Subsequently, experimental validation demonstrated that CH inhibited the phosphorylation of PI3K and AKT in pancreatic tissues, promoted the apoptosis of pancreatic acinar cells, and further alleviated inflammation and histopathological damage to the pancreas in AP rats.
ConclusionApoptosis of pancreatic acinar cells can be enhanced and the inflammatory response can be reduced through the modulation of the PI3K/AKT signaling pathway, resulting in the amelioration of pancreatic disease.

研究目的
本研究旨在采用网络药理学方法分析CH的活性成分及信号通路，并验证CH治疗急性胰腺炎（AP）的分子机制。

材料与方法
首先采用网络药理学与分子对接技术，筛选CH治疗AP的活性成分及其潜在作用靶点。通过逆行向大鼠胰胆管内注射3.5%牛磺胆酸钠，构建急性胰腺炎（AP）大鼠模型。采用组织学检查、酶联免疫吸附试验、蛋白质印迹法（Western blot）及TUNEL染色，探究CH治疗AP的作用通路与分子机制。

结果
网络药理学分析共筛选得到168种活性成分与276个靶点蛋白。此外，与AP相关的靶点共计2060个，其中CH与AP共有的靶点达177个。上述共有靶点（包括STAT3、IL6、MYC、CDKN1A、AKT1、MAPK1、MAPK3、MAPK14、HSP90AA1、HIF1A、ESR1、TP53、FOS及RELA）被鉴定为核心靶点。进一步从基因本体（Gene Ontology, GO）数据库筛选得到5252条条目，从京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）数据库筛选得到186条信号通路。蛋白质相互作用富集分析与网络分析结果显示，CH可显著调控PI3K/AKT信号通路，该通路在程序性细胞死亡中发挥关键作用。分子对接结果证实，核心活性成分与关键靶点间具有较强的结合活性。后续实验验证结果表明，CH可抑制AP大鼠胰腺组织中PI3K与AKT的磷酸化水平，促进胰腺腺泡细胞凋亡，进而减轻AP大鼠的胰腺炎症反应与组织病理学损伤。

结论
通过调控PI3K/AKT信号通路，可增强胰腺腺泡细胞凋亡、减轻炎症反应，从而改善胰腺病变。