文献类型：期刊文献

中文题名：芦银方解热抗炎作用及机制研究

英文题名：Antipyretic and Anti-Inflammatory Effects and Mechanism of Lu Yin(芦银)Formula

作者：杨丽丽[1];刘小瑞[1];海洋[1];杨文[1];李梓慕[1];王斯琦[1];杨瑾瑾[1];汪永锋[2];刘东玲[1,3,4]

第一作者：杨丽丽

机构：[1]甘肃中医药大学药学院,兰州730000;[2]甘肃中医药大学基础医学院,兰州1730000;[3]甘肃中医药大学陇药产业创新研究院,兰州730000;[4]西北中藏药省部共建协同创新中心,兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2024

卷号：40

期号：12

起止页码：26

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：2020年甘肃省高等学校产业支撑计划项目(编号:2020C-36);甘肃省教育厅“双一流”科研重点项目(编号:GSSYLXM-05);杰出青年基金项目(编号:23JRRA1230)。

语种：中文

中文关键词：芦银方;解热抗炎;网络药理学;分子对接;核因子激活的B细胞的κ-轻链增强子;双氧合酶2;丝裂原活化蛋白激酶

外文关键词：Lu Yin(芦银)Formula;Antipyretic and anti-inflammatory;Network pharmacology;Molecular docking;NF-kB;MAPK

摘要：目的:基于网络药理学、分子对接和动物实验探讨芦银方解热、抗炎作用及主要机制。方法:通过TCMSP和BATMAN-TCM数据库得到芦银方的活性成分及靶标蛋白,采用GeneCard、TTD、DrugBank及OMIM数据库获得发热和炎症相关靶点。运用Jvenn在线平台获取药物疾病共有靶点获得芦银方解热抗炎的潜在靶点,构建中药-化合物-潜在靶点-疾病网络图,筛选出芦银方解热抗炎核心成分。通过STRING数据库构建潜在靶点相互作用(PPI)网络图,并通过Degree值筛选出核心靶点,借助DAVID在线平台对核心靶点进行KEGG及GO富集分析,将核心成分与核心靶点运用AutoDock vina 1.5.6软件进行分子对接。构建二甲苯致小鼠耳廓肿胀模型,观察芦银方的抗炎作用;建立干酵母致大鼠发热模型,观察芦银方的解热作用;利用体温监测、HE染色、ELISA法和Western blot法评估芦银方药效并对p-JNK、p-P38、p-NF-κBp65、p-ERK和PTGS2炎症及发热关键蛋白的表达进行考察。结果:网络药理学预测得到芦银方治疗发热、炎症的10个核心成分,包括白桦脂醇和香叶基丙酮等,核心治疗靶点包括PTGS2、NF-κB、MAPK3/1(ERK1/2)、MAPK8(JNK)和MAPK14(p38)等。GO富集分析结果发现发热和炎症反应与MAPK激酶活性的调控、MAPK级联、NF-κB结合及ERK的正调控等相关。KEGG通路富集结果显示,芦银方可能通过调控MAPK、NF-κB及TNF等信号通路发挥解热抗炎作用。分子对接显示,白桦脂醇与JNK和ERK蛋白结合能最高,而槲皮素与PTGS2、P38及NF-κB蛋白均有较高的结合能。动物实验结果表明,与模型对照组相比,芦银方显著抑制小鼠耳肿胀,具有明显的抗炎作用(P<0.05)。干酵母致热实验结果表明,与模型对照组相比,芦银方10.8、21.6 g/kg组大鼠连续时间段内肛温显著下降。组织病理学HE染色显示,模型对照组大鼠下丘脑神经元数量减少、胞质疏松并且胞核皱缩深染,神经纤维结构疏松并可见较多的小空泡,芦银方给药后可见少量的神经元胞质疏松且可见的小空泡变少,病理状况明显改善。与模型对照组相比,芦银方10.8、21.6 g/kg组大鼠下丘脑和血清中致热炎性因子TNF-α、IL-1β、IL-6、PGE2含量显著降低(P<0.01),下丘脑p-P38、p-JNK、p-NF-κBp65、p-ERK、PTGS2蛋白表达下调(P<0.05)。结论:芦银方可能通过白桦脂醇和槲皮素等核心化合物,调控MAPK/NF-кB/PTGS2信号通路,抑制内生致热炎性因子IL-6、IL-1β、TNF-α及PGE2的产生,减轻神经元损伤,发挥解热抗炎作用。
Objective: To investigate the antipyretic and anti-inflammatory effects and the main mechanisms of Lu Yin(芦银) Formula based on network pharmacology, molecular docking, and animal experiments. Methods: The TCMSP and BATMAN-TCM databases were used to obtain the active ingredients and target proteins of Lu Yin Formula. GeneCard, TTD,DrugBank, and OMIM databases were used to obtain the fever and inflammation-related targets. Jvenn online platform was utilized to obtain the potential targets of Lu Yin Formula for antipyretic and anti-inflammatory effects by taking the common targets of drugs and diseases, and the network diagram of traditional Chinese medicine(TCM)-compound-potential target-disease was constructed to screen out the core components of Lu Yin Formula for antipyretic and anti-inflammatory effects. The STRING database was used to construct a network diagram of potential target interactions(PPI),and the core targets were screened out by the Degree value. KEGG and GO enrichment analyses were performed on the core targets by DAVID online platform, and molecular docking between the core components and the core targets was performed using AutoDock vina 1.5.6 software. A xylene-induced auricular swelling model of mice was constructed to observe the anti-inflammatory effect of Lu Yin Formula. A dry yeast-induced fever model of mice was established to observe the antipyretic and anti-inflammatory effects of Lu Yin Formula. Temperature monitoring, HE staining, ELISA,and Western blotting methods were utilized to assess the pharmacological efficacy of Lu Yin Formula and investigate the levels of p-JNK,p-P38,p-NF-κBp65,p-ERK,PTGS2,and key proteins for inflammation and fever. Results: Network pharmacology predicted 10 core components of Lu Yin Formula for the treatment of fever and inflammation, including betulinol and geranylacetone. The core therapeutic targets included PTGS2,NF-κB,MAPK3/1(ERK1/2),MAPK8(JNK),and MAPK14(p38). GO enrichment analysis revealed that fever and inflammatory responses were associated with the regulation of MAPK kinase activity, MAPK cascade, NF-kB binding, and positive regulation of ERK. KEGG pathway enrichment results showed that Lu Yin Formula might exert antipyretic and anti-inflammatory effects through signaling pathways such as MAPK,NF-κB,and TNF. Molecular docking revealed that betulinol had the highest binding energy to JNK and ERK proteins, whereas quercetin had a high binding energy to PTGS2,P38,and NF-κB proteins. Animal experiments showed that compared with the model control group, Lu Yin Formula significantly inhibited ear swelling in mice and had a significant anti-inflammatory effect(P<0.05). The results of the dry yeast thermogenic assay showed that compared with those in the model control group, the anal temperatures of mice in the Lu Yin Formula groups of 10.8 and 21.6 g/kg were decreased significantly over successive time periods. Histopathological HE staining showed that the number of hypothalamic neurons in the model control group was reduced;the cytoplasm was loose, and the nuclei were crumpled and deeply stained. The structure of nerve fibers was loose, and more small vacuoles were observed. After Lu Yin Formula treatment, a small number of neurons with loose cytoplasm and fewer small vacuoles were observed, and the pathological condition was significantly improved. Compared with the model control group, the Lu Yin Formula groups of 10.8 and 21.6 g/kg reduced the contents of the thermogenic inflammatory factors TNF-α,IL-1β,IL-6,and PGE2 in the hypothalamus and serum of mice(P<0.01) and down-regulated the protein expressions of p-P38,p-JNK,p-NF-κBp65,p-ERK,and PTGS2(P<0.05),so as to exert an antipyretic and anti-inflammatory effect. Conclusion: Lu Yin Formula may modulate the MAPK/NF-кB/PTGS2 signaling pathway through core compounds such as betulinol and quercetin, inhibit the production of endogenous thermogenic inflammatory factors IL-6,IL-1β,TNF-α,and PGE2,and attenuate neuronal damage to exert antipyretic and anti-inflammatory effects.