文献类型：期刊文献

中文题名：基于网络药理学与非靶向代谢组学探讨细辛的肝损伤机制

英文题名：Discussion on hepatic damage mechanism of Asari Radix et Rhizoma based on network pharmacology and untargeted metabolomics

作者：鲍慧中[1];朱丽娟[1,2];刘雪枫[1,2];王丽娟[1,2];罗慧英[1,2,3,4]

第一作者：鲍慧中

机构：[1]甘肃中医药大学药学院,甘肃兰州730101;[2]甘肃省中药药理与毒理学重点实验室,甘肃兰州730000;[3]陇药产业创新研究院,甘肃兰州730000;[4]西北中藏药省部共建协同创新中心,甘肃兰州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：49

期号：10

起止页码：2575

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；PubMed;

基金：甘肃省自然科学基金项目(22JR5RA585)。

语种：中文

中文关键词：细辛;肝损伤;网络药理学;非靶向代谢组学

外文关键词：Asari Radix et Rhizoma;hepatic damage;network pharmacology;untargeted metabolomics

摘要：细辛是临床常用解表药,但其毒性限制了其使用。该研究采用网络药理学与代谢组学相结合的方法探讨细辛的肝损伤机制。GEO数据库下载肝损伤相关数据集GSE54257,采用Limma包对数据集GSE54257进行差异表达基因分析。TCMSP、ECTM、TOXNET等数据库筛选细辛有毒成分与靶点基因,与GSE54257差异表达基因相映射,得到细辛肝损伤靶点基因并构建PPI网络。对靶点基因进行GO和KEGG富集分析,结合上游调控miRNA信息绘制“miRNA-靶点基因-信号通路”网络。30只大鼠分为空白组与细辛高、低剂量组,每日给药1次,连续给药28 d后检测肝功能指标和肝脏病理变化。空白组与细辛高剂量组随机取5个肝脏组织样本,采用超高效液相色谱-质谱(UHPLC-MS)技术分析各组样本小分子代谢产物。利用正交偏最小二乘法判别分析(OPLS-DA)筛选差异代谢物,并对差异代谢物进行富集分析、相关性分析和聚类分析,最后利用MetaboAnalyst平台对差异代谢物进行通路富集分析。经分析发现,细辛中有毒成分有14个,对应靶点基因37个,经与GSE54257中差异表达基因映射得到12个与细辛肝脏毒性相关的基因。动物实验结果显示,细辛可以显著升高大鼠肝功能指标,降低自由基清除酶活性,改变肝脏氧化应激水平,诱发脂质过氧化损伤。非靶向代谢组学分析结果显示,与空白组比较,细辛组大鼠肝脏组织中有9个代谢物上调,16个代谢物下调,这25个代谢物相关性较强,聚类良好。通路富集分析显示,这些差异代谢物与细辛12个肝毒性靶点基因主要参与嘌呤代谢,以及缬氨酸、亮氨酸、甘氨酸、丝氨酸、苏氨酸等的生物合成与代谢。该研究证实,细辛致肝损伤作用是多成分、多靶点、多信号通路作用的结果,其机制可能与抑制核苷酸合成,影响蛋白质代谢有关。
Asari Radix et Rhizoma is a common drug for relieving exterior syndrome in clinics,but its toxicity limits its use.In this study,the mechanism of hepatic damage of Asari Radix et Rhizoma was studied by network pharmacology and metabolomics.The hepatic damage-related dataset,namely GSE54257 was downloaded from the GEO database.The Limma package was used to analyze the differentially expressed genes in the dataset GSE54257.Toxic components and target genes of Asari Radix et Rhizoma were screened by TCMSP,ECTM,and TOXNET.The hepatic damage target genes of Asari Radix et Rhizoma were obtained by mapping with the differentially expressed gene of GSE54257,and a PPI network was constructed.GO and KEGG enrichment analysis of target genes were performed,and a "miRNA-target gene-signal pathway" network was drawn with upstream miRNA information.Thirty rats were divided into a blank group,a high-dose Asari Radix et Rhizoma group,and a low-dose Asari Radix et Rhizoma group,which were administered once a day.After continuous administration for 28 days,liver function indexes and liver pathological changes were detected.Five liver tissue samples were randomly collected from the blank group and high-dose Asari Radix et Rhizoma group,and small molecule metabolites were analyzed by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS).The orthogonal partial least squares-discriminant analysis(OPLS-DA) method was used to screen differential metabolites,and enrichment analysis,correlation analysis,and cluster analysis were conducted for differential metabolites.Finally,the MetaboAnalyst platform was used to conduct pathway enrichment analysis for differential metabolites.It was found that there were 14 toxic components in Asari Radix et Rhizoma,corresponding to 37 target genes,and 12 genes related to liver toxicity of Asari Radix et Rhizoma were obtained by mapping to differentially expressed genes of GSE54257.The animal test results showed that Asari Radix et Rhizoma could significantly increase the liver function index,reduce the activity of the free radical scavenging enzyme,change the liver oxidative stress level,and induce lipid peroxidation damage in rats.The results of untargeted metabolomics analysis showed that compared with the blank group,nine metabolites were up-regulated,and 16 metabolites were down-regulated in the liver tissue of the Asari Radix et Rhizoma group.These 25 metabolites had strong correlations and good clustering.Pathway enrichment analysis showed that these differential metabolites and the 12 hepatotoxic target genes of Asari Radix et Rhizoma were mainly involved in purine metabolism,as well as the biosynthesis and metabolism of valine,leucine,glycine,serine,and threonine.The study confirmed that the hepatica damage effect of Asari Radix et Rhizoma was the result of multi-component,multi-target,and multi-signaling pathways,and its mechanism may be related to inhibiting nucleotide synthesis and affecting protein metabolism.