海南医科大学学报2025，Vol.31Issue(12)：938-953,16.DOI:10.13210/j.cnki.jhmu.20241106.001
基于网络药理学、分子对接和细胞实验探讨高粱泡叶醇提物抗炎的作用机制
Exploring the anti-inflammatory mechanism of action of Rubus lambertianus leaf ethanol extract based on network pharmacology,molecular docking and cellular experiments
摘要
Abstract
Objective:To explore the anti-inflammatory efficacy and mechanism of action of Rubus lambertianus leaf ethanol extract(AELR)based on network pharmacology,molecular docking and cellular experimental validation.Methods:The chemical composition of AELR was analyzed by ultra-high performance liquid chromatography coupled with high-resolution quadrupole or-bit trap mass spectrometry(UHPLC-Q-Exactive HF/MS).The AELR chemistry was screened by SwissADME platform and then the SwissTargetPrediction database was used to retrieve potential targets;OMIM,GeneCards,and TTD databases were used to search for disease targets;Venny2.1 platform was used to obtain"component-disease"intersection targets and construct protein-protein interaction(PPI)networks;GO function and KEGG signaling pathway enrichment analysis of intersecting targets were conducted using the DAVID database;Molecular docking validation of key active ingredients and core targets was performed using software such as AutoDock;Finally,an in vitro inflammation model was constructed using LPS-induced RAW264.7 cells.The levels of cell supernatant-associated inflammatory factors were detected by Griess,ELISA,and flow cytometry,and cell-associated protein mRNA and signaling pathway protein expression were detected by RT-PCR and Western blot techniques.Results:A total of 56 compounds were identified by UHPLC-Q-Exactive HF/MS;21 AELR active ingredients and 419 targets were obtained by network pharmacological screening,with 1 923 inflammatory disease targets and 187 intersecting targets,among which IL-1B,AKT1,STAT3,EGFR,PTGS2 were the core targets,and the molecular docking results with the active ingredients of stigmasterol,lignoceroside,isoxanthin,brown cornflower,cloverberry Xanthin,brown cornflower,alfalfa active ingredient molec-ular docking results were good;GO and KEGG analyses revealed that AELR anti-inflammatory may be transduced through PI3K/AKT,MAPK,and NF-κB signaling pathways.In vitro cellular experiments showed that AELR significantly reduced NO,IL-6,TNF-αand intracellular ROS levels in cell supernatants compared with the model group(P<0.01),significantly reduced cellular COX-2、IL-6、TNF-α、IL-1β mRNA expression reduction(P<0.01),and significantly down-regulated p-p65/p65、p-IκBα/IκBα、p-JNK/JNK、p-ERK/ERK、p-Akt/Akt protein expression(P<0.01).Conclusion:AELR has anti-inflammatory effects,and its anti-inflammatory mechanism of action may be inhibition of the protein expression of NF-κB,MAPK and Akt signaling pathways through multiple components and targets.关键词
高粱泡叶/抗炎/网络药理学/UHPLC-Q-Exactive HF/MS/作用机制Key words
Rubus lambertianus leaf/Anti-inflammatory/Network pharmacology/UHPLC-Q-Exactive HF/MS/Mecha-nism of action分类
医药卫生引用本文复制引用
田孟斌,吴静澜,李旭东,韩伟,韦美燕,聂琴,陈汝玲..基于网络药理学、分子对接和细胞实验探讨高粱泡叶醇提物抗炎的作用机制[J].海南医科大学学报,2025,31(12):938-953,16.基金项目
This study was supported by Traditional Chinese Medicine and Ethnic Medicine Science and Technology Project of Guizhou Provincial Administration of Traditional Chinese Medicine(QZYY-2022-011)and Research Center for Medicinal Polymer Materials at Guizhou University of Traditional Chinese Medicine(Guizhong Medicine Party Office[2019]No.70) 贵州省中医药管理局中医药、民族医药科学技术课题(QZYY-2022-011) （QZYY-2022-011）
贵州中医药大学药用高分子材料研究中心(贵中医党办发[2019]70号) （贵中医党办发[2019]70号）
