基于网络药理学分析火棘果实提取物的抗炎作用及实验研究  

Analysis of the anti-inflammatory effect and experimental study of Pyracantha fruit extracts based on network pharmacology

作　　者：朱亮 伍明江[2] 邱佳[2] 范芳芳 梁冰 ZHU Liang;WU Mingjiang;QIU Jia;FAN Fangfang;LIANG Bing(Department of Pharmacology,School of Basic Medicine,Guizhou Medical University,Guiyang 561113,Guizhou,China;Department of Pharmacy,Zunyi Medical and Pharmaceutical College,Zunyi 563006,Guizhou China)

机构地区：[1]贵州医科大学基础医学院药理学教研室,贵州贵阳561113 [2]遵义医药高等专科学校药学系,贵州遵义563006

出　　处：《贵州医科大学学报》2025年第3期346-360,共15页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82460781);“全国中药资源普查”专项(国中医药办科技函[2018]132号,国中医药办科技函[2019]186号);遵义市科技合作计划项目(遵市科合HZ字[2019]204号)。

摘　　要：目的基于网络药理学预测火棘果实提取物的活性成分,并验证其抗炎作用。方法通过数据库筛选火棘果实提取物与炎症相关的靶点,并分析药物靶点与疾病靶点的交集;制备火棘果实提取物,使用质谱分析其化学成分;通过小鼠耳肿胀模型比较耳肿胀度,构建大鼠足跖肿胀模型,检测大鼠的足跖肿胀率及炎症因子[白细胞介素1β(interleukin-1β,IL-1β)、IL-6、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)和前列腺素E_(2)(prostaglandin E_(2),PGE_(2))]变化,计算其胸腺(脾脏)指数,从而评价火棘果实提取物的抗炎作用;构建小鼠单核巨噬细胞白血病细胞(RAW264.7)炎症模型,检测火棘果实提取物对细胞存活率、细胞上清液炎症因子NO、IL-1β、IL-6、TNF-α及PGE_(2)的浓度及其细胞蛋白炎症因子[IL-1β、IL-6、TNF-α、环氧化酶(cyclooxygenase-2,COX-2)]表达水平的影响;运用网络药理学方法进一步预测火棘果实提取物的作用分子靶点。结果预测了3种抗炎成分,识别出130个药物与疾病交集靶点;质谱分析鉴定火棘果实提取物中的11种化合物,原花青素B1、槲皮素、儿茶素与预测的抗炎成分一致;在二甲苯诱导的小鼠耳朵肿胀实验中,火棘低、中、高剂量组耳肿胀度低于对照组(P<0.05);在角叉菜胶诱导的大鼠足跖肿胀实验中,火棘各高剂量组足跖肿胀度低于对照组(P<0.05),火棘低、中、高剂量组IL-1β、IL-6和TNF-α表达降低(P<0.05);火棘中、高剂量组的PGE_(2)表达减少(P<0.05),火棘低、中、高剂量组胸腺和脾脏指数升高(P<0.05);RAW264.7细胞炎症模型实验中,细胞活性结果显示,火棘果实提取物的半抑制浓度(IC_(50))为242.1 mg/L,火棘各剂量组细胞上清液和蛋白中炎症因子IL-1β、IL-6、TNF-α的表达减少(P<0.05),NO、PGE_(2)、COX-2表达降低(P<0.05);蛋白质互作网络(protein-protein interaction,PPI)分析识别出33个核心靶点,基因本体论(gene ontology,GO)和Objective This investigation focuses on predicting the active components of Pyracantha fruit extracts via network pharmacology and experimentally confirming its anti-inflammatory effect.Methods The database was used to screen the targets related to the Pyracantha fruit extracts associated with inflammation,and analyze the overlap between drug targets and disease targets.The chemical composition of Pyracantha fruit extracts were prepared and analyzed using mass spectrometry.The degree of ear swelling was compared among mouse ear swelling models and the rat paw swelling model was established to measure paw swelling and inflammatory factors:interleukin 1β(IL-1β),interleukin-6(IL-6),tumor necrosis factorα(TNF-α),and prostaglandin E_(2)(PGE_(2)).The thymus and spleen indices were calculated to evaluate the anti-inflammatory effects of the Pyracantha fruit extracts.The mouse monocyte macrophage leukemia cell(RAW264.7)inflammation model was established to detect the effects of the Pyracantha fruit extracts on cell viability and the concentrations of inflammatory factors in cell supernatant,such as nitric oxide(NO),IL-1β,IL-6,TNF-α,PGE_(2),and the expression levels of cell protein inflammatory factors IL-1β,IL-6,TNF-α,and cyclooxygenase(COX-2).And network pharmacology methods were used to further predict the molecular targets of the Pyracantha fruit extracts.Results Three anti-inflammatory components were predicted,leading to the identification of 130 drug-disease intersection targets.Additionally,mass spectrometry analysis revealed 11 compounds in the Pyracantha fruit extracts,including procyanidin B1,quercetin,and catechin,which matched the predicted anti-inflammatory components;in the xylene-induced mouse ear swelling experiment,the degree of ear swelling in the low,medium,and high dose groups of Pyracantha was lower than that of the control group(P<0.05);In the carrageenan-induced rat paw swelling experiment,the degree of paw swelling in the high dose group of Pyracantha was significantly lower than that of t

关 键 词：火棘果实提取物 质谱分析 抗炎作用 网络药理学 分子对接 

分 类 号：R285[医药卫生—中药学]