基于LC/GC-MS和网络药理学探讨大黄-桃仁药对逐瘀泻热的作用机制  

Active ingredients and mechanism of Rhubarb-Peach kernel pair in removing blood stasis and purging heat based GC-MS and network pharmacology

作　　者：黄建萍 井亚江 王七龙 邹远荣 夏泽冰 王红艳 彭亮 张岗 高静 颜永刚 HUANG Jian-ping;JING Ya-jiang;WANG Qi-long;ZOU Yuan-rong;XIA Ze-bing;WANG Hong-yan;PENG Liang;ZHANG Gang;GAO Jing;YAN Yong-gang(Shaanxi Engineering Research Center for Application and Development of Chinese Medicine in Qinling Mountains/Key Laboratory of Qin Medicine Research and Development,Shaanxi University of Chinese Medicine,Xianyang Shaanxi 712046)

机构地区：[1]陕西中医药大学药学院/陕西省秦岭中草药应用开发工程技术研究中心/“秦药”研发重点实验室,陕西咸阳712046

出　　处：《中南药学》2024年第4期892-901,共10页Central South Pharmacy

基　　金：国家自然科学基金项目(No.81973592);陕西省中医药管理局项目(No.2021-GJ-JC004)。

摘　　要：目的 分析大黄-桃仁药对水煎液的化学成分,探讨其逐瘀泻热的药效物质及作用机制。方法 采用UPLC-TOF-MS、GC-MS对大黄-桃仁药对水煎液进行成分分析,结合网络药理学方法构建大黄-桃仁药对逐瘀泻热的“疾病-成分-靶点”网络,筛选目标信号通路。采用间隔24 h两次大鼠尾静脉注射内毒素脂多糖复制“瘀热”大鼠模型,通过大鼠体温变化,血液流变、凝血四项(TT、PT、APTT、FIB)、炎症因子水平、氧化应激水平等的测定对大黄-桃仁药对逐瘀泻热进行药效评价。通过分子对接技术、实时荧光定量聚合酶链式反应(RT-q PCR)对预测的核心靶点进行验证。结果 从大黄-桃仁药对水煎液中鉴定得到化合物64个,网络药理学分析结果表明大黄-桃仁药对中芦荟大黄素、圣草酚、表没食子儿茶素没食子酸酯、原花青素等核心成分,通过作用于AKT1、PTGS2、SRC、IL1B、TNF等核心靶点,调控VEGF、MAPK、IL-17、TNF等通路达到逐瘀泻热的目的。动物实验结果表明,大黄-桃仁药对给药后,模型大鼠的氧化应激(NO、SOD、MDA)、凝血四项、血液流变、体温等指标均向正常回调,大黄-桃仁药对能够降低炎症因子IL-1β、IL-17、TNF-α含量以及核心靶点AKT1、SRC、MYC、EGFR、PTGS2、TP53 mRNA表达。结论 圣草酚、芦荟大黄素、原花青素、没食子酸儿茶素、表没食子儿茶素没食子酸酯等化合物为大黄-桃仁药对逐瘀泻热的药效物质。大黄-桃仁药对通过多靶点,多通路调控机体炎症、氧化应激、血管生成、血管内皮等方面发挥逐瘀泻热作用。Objective To analyze the chemical compositions of the decoction of Rhubarb-Peach kernel pair,and to determine the pharmacodynamic substances and mechanism of action in removing blood stasis and purging heat.Methods UPLC-TOF-MS and GC-MS were used to analyze the composition of Rhubarb-Peach kernel pair decoction.Network pharmacology was used to establish the“disease-component-target”network of Rhubarb-Peach kernel pair and screen the target signaling pathways.The rat model of stasis-heat was replicated by tail vein injection of endotoxin lipopolysaccharide twice at an interval of 24 hours.The efficacy of Rhubarb-Peach kernel pair was evaluated with changes in body temperature,blood rheology,four coagulation items(TT,PT,APTT,and FIB),inflammatory factor levels,and oxidative stress levels in rats.The predicted core targets were verified by molecular docking and real-time fluorescence quantitative polymerase chain reaction.Results Totally 64 compounds of Rhubarb-Peach kernel pair decoction were identified.Network pharmacological indicated that the core components of Rhubarb-Peach kernel pair,such as aloe-emodin,eriodictyol,epigallocatechin gallate,and procyanidin modulated the pathways of VEGF,MAPK,IL-17,and TNF to remove blood stasis and purge heat through the core targets,such as AKT1,PTGS2,SRC,IL1B and TNF.Docking between the active ingredients and core targets molecules were favorable.The animal experiments showed that after the Rhubarb-Peach kernel pair treatment,the oxidative stress(NO,SOD,and MDA),four coagulation items,blood rheology,body temperature and other indicators of the blood of stasis-heat model rats all returned to normal levels.Rhubarb-Peach kernel pair reduced the contents of inflammatory factors IL-1β,IL-17,and TNF-α,as well as the expression of core targets,namely AKT1,SRC,MYC,EGFR,PTGS2,and TP53 mRNA.Conclusion Compounds such as eriodictyol,aloe-emodin,procyanidin,catechin gallte,and epigallocatechin gallate are the material basis for Rhubarb-Peach kernel pair to remove blood stasis and p

关 键 词：大黄-桃仁药对 逐瘀泻热 UPLC-TOF-MS GC-MS 网络药理学 药效学实验 

分 类 号：R285[医药卫生—中药学] R286[医药卫生—中医学]