“量-权-证”网络毒理学的提出与应用:以八角莲醇提液致肝毒性为例

该研究首次提出"量-权-证"网络毒理学的研究方法,通过将毒性成分含量及成分作用靶点的频次赋予靶点权重,将传统定性网络转为定量网络,不断提高获取数据的信度,为系统性评价中药安全性及毒理学研究提供研究思路。首先,给予大鼠灌胃八角莲50%醇提物,基于血清药物化学辨识其中成分。随后从SwissTargetPrediction、PharmMapper等数据库获取成分靶点,以成分相对含量和钓靶频次赋予靶点权重,Comparative Toxicogenomics Database(CTD)、GeneCards等数据库预测肝毒性靶点,通过STRING数据库进行蛋白质互作分析及通路富集分析。最后,构建"毒性成分-加权靶点-效应通路"定量网络。八角莲体内外成分分析共筛选得到鬼臼毒素、鬼臼毒酮、去氧鬼臼毒素、6-甲氧基鬼臼毒素等11个潜在毒性物质,涉及肝毒性作用靶点106个,加权靶点涉及Cdk2、Egfr、Cyp2c9等65个。京都基因与基因组百科全书数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析表明其可能通过作用于PI3K-AKT、MAPK、Ras等信号通路呈现炎症反应、氧化应激、细胞凋亡、介导细胞色素P450(CYP450)酶系发挥肝毒性作用,然传统方法显示AKT1、Alb、Stat3等51个靶点可能通过介导炎症作用、细胞增殖等导致肝损伤,可知从不同层面获取的毒性机制有较大差异。综上,该研究提出并系统性应用"量-权-证网络毒理学"于八角莲致大鼠肝毒性机制研究,证实其广泛应用于中药毒理学评价的可行性,进一步完善中药安全性系统评价的思路。

Mechanism of hepatotoxicity induced by ethanol extract of Dysosma versipellis based on"quantity-weight-evidence"network toxicology

In this study,the toxicological/pharmacological research method of"quantity-weight-evidence"network was first proposed and practiced to supplement the existing methodology of network toxicology.We transformed the traditional qualitative network into a quantitative network in this study by attributing weights to toxic component content and target frequency,which improved the reliability of data and provided a research idea for the systematic safety evaluation and toxicological research of Chinese medicinal herbs.Firstly,50%ethanol extract of Dysosma versipellis(DV)was administrated to rats via gavage and the potential hepatotoxic components were identified by serum pharmacochemistry.Then,the component targets were obtained from SwissTargetPrediction,PharmMapper and other online databases,and the target weights were given according to the relative content of components and target fishing frequency.Meanwhile,the targets of hepatotoxicity were predicted from online databases such as Comparative Toxicology Database(CTD)and GeneCards.Subsequently,protein-protein interaction analysis and KEGG pathway enrichment were performed with the STRING database.Finally,the quantitative network of"toxic components-weighted targets-pathways"was constructed.Eleven potential toxic compounds were predicted,including podophyllotoxin,podophyllotoxone,deoxypodophyllotoxin,and 6-methoxypodophyllotoxin.A total of 106 hepatotoxic targets and 65 weighted targets(e.g.,Cdk2,Egfr,and Cyp2 c9)were identified.The results of Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment showed that these targets could act on PI3 K-AKT,MAPK,and Ras signaling pathways to play a role in inflammatory response and oxidative stress.However,traditional network toxicology showed that 51 targets such as AKT1,Alb,and Stat3 may lead to hepatotoxicity by mediating inflammation and cell proliferation.In conclusion,we proposed"quantity-weight-evidence"network toxicology in this study and used it to study the mechanism of DV-induced hepatotoxicity in rats.This study confirms the feasibility of this new methodology in toxicological evaluation and further improves the systematic evaluation of the safety of Chinese medicinal herbs.