附子生物碱类成分分析及其抗肿瘤机制初探

Analysis of Alkaloids in Aconiti Lateralis Radix Praeparata and Prediction of Their Mechanism in the Treatment of Cancer

目的利用超高效液相色谱-三重四极杆质谱联用法(UPLC-MS/MS)测定附子水煎液中生物碱成分,并运用网络药理学探究其抗肿瘤的多成分、多靶点、多途径作用机制,为附子抗肿瘤的物质基础研究提供参考。方法使用UPLC-MS/MS技术定量分析附子水煎液中12种生物碱含量。借助Swiss Target Prediction、Gene Cards平台预测附子生物碱成分与肿瘤的疾病靶点,用Cytoscape软件构建相关作用网络,并对核心基因进行GO分析与KEGG通路富集。结果附子水煎液中单酯型生物碱含量最高,约占59.07%,醇胺型生物碱含量次之,约为29.48%。网络药理学推测附子生物碱抗肿瘤关键靶点为甲硫腺苷磷酸化酶(MTAP)、ATP结合盒B亚家族成员1(ABCB1)基因,并与炎症反应、磷脂酰肌醇、缺氧诱导因子、雌激素、中心碳代谢等紧密相关。结论该方法能同时定量分析附子中12种生物碱成分,简便可行、准确可靠,同时网络药理学结果可为附子临床抗肿瘤应用提供依据,为后期分子生物学研究奠定基础。

OBJECTIVE To quantify the alkaloids in aqueous extract of Aconiti Lateralis Radix Praeparata(Fuzi in Chinese)by ultra performance liquid chromatography coupled with triple quadrupole mass spectrometry(UPLC-MS/MS),meanwhile investigate the anti-cancer mechanism of these alkaloids by network pharmacology.This article aimed to provide a reference for the material basis of anticancer effect of Fuzi.METHODS UPLC-MS/MS was used to analyze 12 alkaloids in Fuzi aqueous extract.The alkaloids and disease targets were predicted by Swiss Target Prediction and Gene Cards platform.The related function network was constructed by Cytoscape software.Finally,the key gene was analyzed by GO and KEGG pathway enrichment analysis.RESULTS Monoester alkaloids accounted for the highest proportion(59.07%),followed by alcohol amine alkaloids(29.48%).Network pharmacology speculated that MTAP(methylthioadenosine phosphorylase)and ABCB1(ATP binding cassette subfamily B member 1)genes were the key targets for the anticancer efficacies of active alkaloids.The key targets were closely related to inflammatory response,phosphatidylinositol-mediated signaling,HIF-1 signaling pathway,estrogen signaling pathway and central carbon metabolism in cancer.CONCLUSION The developed method can determine 12 kinds of alkaloids in Fuzi simultaneously.The method is simple,feasible,accurate and reliable,which can provide the basis for clinical anticancer application and laid the foundation for further molecular biological research of Fuzi.