基于网络药理学及细胞实验的乳香-没药功效成分抗炎机制研究  

Ant i-inflammatory mechanism of active components in Olibanum and Myrrha based on network pharmacology and cell experiments

作　　者：赵子樟 李佳晌 宿树兰[1] 朱悦[1] 钱大玮[1] 段金廒[1] ZHAO Zi-zhang;LI Jia-shang;SU Shu-lan;ZHU Yue;QIAN Da-wei;DUAN Jin-ao(National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine,Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学江苏省中药资源产业化过程协同创新中心,中药资源产业化与方剂创新药物国家地方联合工程研究中心,江苏南京210023

出　　处：《中国中药杂志》2021年第21期5674-5682,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30973885)。

摘　　要：3-羰基-甘遂-8,24-二烯-21-羧酸(KTDA)和2-甲氧基-5-乙酰基-呋喃吉玛烷-1(10)-烯-6-酮(FSA)分别为从乳香和没药中分离得到的萜类化合物,在制备过程中具有得率高且易于结晶的特点。该研究基于网络药理学和细胞活性验证探讨KTDA和FSA的网络调控靶标及抗炎作用机制。首先采用Swiss ADME服务器对活性成分进行类药性预测;通过Swiss Target Prediction和Pharmmapper数据库对活性成分进行靶点预测;使用TTD、DrugBank和Gene Cards数据库检索炎症相关基因,进而得到活性成分的炎症相关靶点;通过STRING数据库分别对KTDA和FSA的炎症靶点进行蛋白互作分析,使用Cytoscape软件对互作结果进行拓扑分析并构建蛋白互作网络图;通过DAVID数据库对活性成分的炎症靶点进行GO功能富集分析和KEGG通路富集分析,构建"活性成分-靶点-通路"网络图。最后采用脂多糖(LPS)诱导的RAW264.7细胞炎症模型,分别采用不同浓度的KTDA和FSA处理,检测一氧化氮(NO)浓度、相关蛋白和m RNA的表达水平。结果表明,KTDA和FSA均显示出较好的类药性;KTDA和FSA共筛选出157和142个炎症相关靶点;PPI网络分析显示MAPK1、AKT1、MAPK8、PIK3CA、PIK3R1、EGFR等蛋白可能为其发挥抗炎作用的关键蛋白;KEGG和GO-BP富集得到PI3K/AKT和MAPK信号通路。细胞实验结果表明,KTDA和FSA可通过抑制NO生成、降低JNK、p38、AKT蛋白的磷酸化水平、降低白细胞介素(interleukin,IL)-1β、IL-6的m RNA表达发挥抗炎作用,同时FSA还可抑制ERK磷酸化。研究结果提示KTDA和FSA具有显著的抗炎活性,为乳香-没药的深入研究与开发利用提供了科学依据和重要支撑。Two terpenes, 3-keto-tirucalla-8,24-dien-21-oic acid(KTDA) and 2-methoxy-5-acetoxy-furanogermacr-1(10)-en-6-one(FSA), are isolated from Olibanum and Myrrha respectively, which are characterized by high yield and easy crystallization during the preparation. The present study explored the regulatory targets and anti-inflammatory mechanism of KTDA and FSA based on network pharmacology and cell viability assay. First, the drug-likeness of KTDA and FSA was predicted by Swiss ADME. The target prediction of active components was carried out by Swiss Target Prediction and Pharmmapper. TTD, Drug Bank, and Gene Cards were searched for inflammation-related target genes of KTDA and FSA. Protein-protein interaction(PPI) analysis was performed on the inflammatory targets of KTDA and FSA by STRING, and Cytoscape was used to conduct topological analysis of the interaction results and construct the PPI network. GO function and KEGG pathway enrichment analyses of inflammatory targets of KTDA and FSA were carried out by DAVID, and a " component-target-pathway" network was constructed. Finally, lipopolysaccharide(LPS)-induced RAW264. 7 cells were treated with KTDA and FSA at different concentrations, and nitric oxide(NO) concentration and protein and m RNA expression levels were detected. The results showed that both KTDA and FSA showed good drug-likeness. A total of 157 and 142 inflammation-related targets of KTDA and FSA were screened out. PPI network analysis showed that MAPK1, AKT1, MAPK8, PIK3 CA,PIK3 R1, EGFR, etc. might be the key proteins for the anti-inflammatory effect. PI3 K/AKT and MAPK signaling pathways were obtained by KEGG and GO-BP enrichment. Cell experiment results showed that KTDA and FSA could exert anti-inflammatory effects by inhibiting NO production, reducing the phosphorylation levels of JNK, p38, and AKT proteins, and down-regulating the m RNA expression of interleukin(IL)-1β and IL-6. Meanwhile, FSA could also inhibit ERK phosphorylation. The results indicated that KTDA and FSA had significant anti-inflam

关 键 词：乳香 没药 抗炎机制 网络药理学 

分 类 号：R285[医药卫生—中药学]