| 徐冬梅,陈学智,余玉宏,等.畲药小香勾香豆素类成分RP-HPLC含量测定及抗炎靶点预测[J].浙江中医药大学学报,2023,47(6):579-586. |
| 畲药小香勾香豆素类成分RP-HPLC含量测定及抗炎靶点预测 |
| Determination of Coumarin Ingredients in Xiaoxianggou of She Herb by RP-HPLC and Prediction of the Corresponding Anti-inflammatory Targets |
| DOI：10.16466/j.issn1005-5509.2023.06.001 |
| 中文关键词: RP-HPLC 畲药小香勾 补骨脂素 佛手柑内酯 抗炎作用 网络药理学 |
| 英文关键词: RP-HPLC Xiaoxianggou of She herb psoralen bergapten anti-inflammatory effect network pharmacology |
| 基金项目:浙江省中医药传承创新项目(2023ZF205)；杭州市农业与社会发展科研项目(202204B10) |
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| 中文摘要: |
| [目的] 以反相高效液相色谱(reverse phase high-performance liquid chromatography，RP-HPLC)法测定畲药小香勾香豆素类成分补骨脂素和佛手柑内酯的含量，并结合网络药理学和动物实验验证小香勾的抗炎作用。[方法] 色谱柱为Waters C18柱(4.6 mm×250 mm，5.0 μm)，流动相A为乙腈，流动相B为0.1%甲酸溶液(体积比)，梯度洗脱，流速为0.8 mL·min-1，补骨脂素检测波长245 nm，佛手柑内酯320 nm，柱温25 ℃。利用网络药理学平台收集小香勾香豆素类成分及抗炎相关靶点，利用 Cytoscape 3.8.2软件建立成分-靶点网络，借助网络拓扑分析插件(Cytoscape network centrality analysis，CytoNCA)筛选抗炎核心靶点，进一步采用Metascape软件进行京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomics，KEGG)通路富集分析。通过二甲苯致小鼠耳肿胀法进一步验证小香勾的抗炎作用。[结果] 补骨脂素和佛手柑内酯的质量浓度分别在0.51～102.00 μg·mL-1(r=0.9996)和0.24～48.00 μg·mL-1(r=0.9998)时与峰面积呈良好的线性关系。通过筛选获得小香勾补骨脂素和佛手柑内酯的相关基因57个，抗炎靶点2 148个，交集靶点31个。KEGG 通路富集分析发现，这些靶点主要涉及化学致癌作用-活性氧、丝裂原活化蛋白激酶(mitogen-activated protein kinase，MAPK)信号通路、癌症途径、催乳素信号通路和叶酸生物合成等信号通路。与模型对照组比较，小香勾高剂量组(400 mg·kg-1)能显著抑制二甲苯所致小鼠耳肿胀(P<0.01)，显示小香勾具有良好的抗炎作用。[结论] 建立的RP-HPLC法操作简单，灵敏度高，可用于小香勾质量控制。小香勾香豆素类成分通过多靶点多通路表现出良好的抗炎作用，为后续深入探讨小香勾抗炎机制提供了有利的科学依据。 |
| 英文摘要: |
| [Objective] To simultaneously determinate coumarin ingredients psoralen and bergapten in Xiaoxianggou(Radix et Chulis Fici Angustifoliae) of She herb by reverse phase high-performance liquid chromatography(RP-HPLC)， and validate the anti-inflammatory effect of Xiaoxianggou with network pharmacology and animal experiment. [Methods] The RP-HPLC system consisted of a Waters C18 column(4.6 mm×250 mm， 5.0 μm) by gradient elution using a mixture of acetonitrile-0.1% HAC as the mobile phase at a flow rate of 0.8 mL·min-1. The UV detection wavelengths of psoralen and bergapten were monitored at 245 nm and 320 nm respectively and the column temperature was 25 ℃. The targets of Xiaoxianggou ingredients and anti-inflammatory genes were collected using network pharmacology platform. The herb ingredient-target network was constructed using Cytoscape 3.8.2 software and the core anti-inflammatory targets were further screened by Cytoscape network centrality analysis (CytoNCA). The Kyoto Encyclopedia of Genes and Genomics(KEGG) pathway enrichment analysis of the targets were analyzed with Metascape software. Dimethylbenzene-induced auricle edema in mice was respectively performed to investigate the anti-inflammatory effect of Xiaoxianggou. [Results] There was good linearity and the response ranges of psoralen and bergapten in 0.51~102.00 μg·mL-1(r=0.9996)， 0.24~48.00 μg·mL-1(r=0.9998)， respectively. A total of 57 potential targets of psoralen and bergapten， 2 148 inflammatory genes and 31 intersection targets were screened. The targets were involved in chemical carcinogenesis-reactive oxygen species， mitogen-activated protein kinase(MAPK) signaling pathway， pathways in cancer， prolactin signaling pathway and folate biosynthesis. Compared with model control group， the high dose group of Xiaoxianggou(400 mg·kg-1) could significantly inhibit the ear swelling induced by dimethylbenzene in mice(P<0.01)， indicating that Xiaoxianggou had good anti-inflammatory effect. [Conclusion] The method was simple， sensitive and rapid， and could be used for simultaneous determination of psoralen and bergapten in Xiaoxianggou. Coumarin ingredients of Xiaoxianggou existed good anti-inflammatory effect through multi-target and multi-pathway. The studies provided a scientific basis for future exploring the anti-inflammatory mechanism of Xiaoxianggou. |
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