基于网络药理学探析5种经典方剂治疗肝细胞癌的作用机制

程秋骆; 刘柳; 白长川; 丛庆伟; 朱英

doi:10.3969/j.issn.1001-5256.2021.08.020

基于网络药理学探析5种经典方剂治疗肝细胞癌的作用机制

DOI: 10.3969/j.issn.1001-5256.2021.08.020

程秋骆^{1, 2},
刘柳^{1, 2},
白长川³,
丛庆伟^{1, 2},
朱英^{1, 2, ,}

1.
大连医科大学附属第一医院中西医结合肝病中心, 辽宁大连 116011
2.
大连医科大学中西医结合学院, 辽宁大连 116044
3.
大连市中医医院中医科, 辽宁大连 116013

基金项目:

国家自然科学基金 (81673728)

利益冲突声明: 本文不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。

作者贡献声明: 程秋骆负责文章撰写, 文献查找与整理, 并撰写初稿; 刘柳、丛庆伟参与文章网络药理学部分数据查找并检索相关文献; 白长川负责中医思路部分指导; 朱英负责拟定写作思路, 指导撰写文章并最后定稿。

详细信息

通信作者:
朱英, zhuyingsh52@126.com

中图分类号: R735.7
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出版历程
- 收稿日期: 2021-01-10
- 录用日期: 2021-03-10
- 出版日期: 2021-08-20

Mechanism of action of five classic prescriptions in treatment of hepatocellular carcinoma based on network pharmacology

Qiuluo CHENG^{1, 2},
Liu LIU^{1, 2},
Changchuan BAI³,
Qingwei CONG^{1, 2},
Ying ZHU^{1, 2
, ,}

1.
Liver Disease Center for Combined TCM and Western Medicine, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, China
2.
Institute (College) of Integrative Medicine, Dalian Medical University, Dalian, Liaoning 116044, China
3.
Department of TCM Dalian Hospital of Traditional Chinese Medicine, Dalian, Liaoning 116013, China

Research funding:

National Natural Science Foundation of China (81673728)

摘要

摘要: 目的运用网络药理学及分子对接技术探究逍遥散合四君子汤、茵陈蒿汤、龙胆泻肝汤合下瘀血汤、五皮饮合四君子汤、一贯煎治疗肝细胞癌的作用机制。方法运用TCMSP、TCMID、BATMAN-TCM、TCM-MESH等数据库筛选中药有效成分并预测其作用靶点, 通过TTD、Drugbank、Disgenet、Liverome、OncoDB.HCC、GEO等数据库挖掘肝癌相关靶点; 将药物和疾病靶点进行映射获得交集靶点, 通过可视化软件Cytoscape3.7.1构建核心成分-交集靶点网络以及蛋白相互作用网络, 筛选核心成分及关键基因并在GEPIA数据库进行生存分析, 将筛选出的活性成分和关键基因导入DockThor在线网站进行分子对接。此外, 利用David数据库对交集靶点进行基因本体(GO)富集分析以及基因组百科全书(KEGG)通路分析。结果经过筛选去重获得5种经典方剂活性成分分别为110、19、154、121、51个, 药物作用靶点分别为7426、1435、9544、6619、2427个。筛选去重后获得肝癌疾病靶点4001个。5种方剂经典方剂的成分靶点与肝癌疾病靶点的交集靶点分别为260、169、276、242、192个, 共同Hub基因在GEPIA在线网站生存分析结果为PIK3CA、SRC、MAPK1、MAPK3(P值均＜0.05)、AKT1(P＞0.05)。5种方剂共同活性成分是槲皮素, 其中3种经典方剂(逍遥散合四君子汤、龙胆泻肝汤合下瘀血汤、五皮饮合四君子汤)的共同活性成分是异补骨脂黄酮、Kanzonol W。分子对接结果从总体趋势来看3种成分与PIK3CA、SRC结合能力相对较好。GO富集分析显示这些靶点共同参与对药物的反应、蛋白质磷酸化、炎症反应、血管生成等生物学过程。KEGG富集分析显示共同参与的通路为癌症通路、PI3K-AKT通路、MAPK通路、Ras通路、HIF-1通路、乙型肝炎通路、丙型肝炎通路。结论槲皮素、异补骨脂黄酮和Kanzonol W抗肝癌具有多靶点、多通路的潜在作用机制。
- 癌, 肝细胞 /
- 网络药理学 /
- 信号传导
Abstract: Objective To investigate the mechanism of action of Xiaoyao powder combined with Sijunzi decoction, Artemisia capillaris Thunb. decoction, Longdan Xiegan decoction combined with Xiayuxue decoction, Wupi decoction combined with Sijunzi decoction, and Yiguan decoction in the treatment of hepatocellular carcinoma (HCC) based on network pharmacology and molecular docking. Methods Databases including TCMSP, TCMID, BATMAN-TCM, and TCM-MESH were used to screen out effective components and predict their targets, and databases including TTD, Drugbank, Disgenet, Liverome, OncoDB.HCC, and GEO were used to investigate HCC-related targets. The drug and disease targets were mapped to obtain the intersecting targets, and the visualization software Cytoscape 3.7.1 was used to construct the core component-intersecting target network and the protein-protein interaction (PPI) network. The core components and key genes were screened out and a survival analysis was performed in the GEPIA database. The active components and key genes screened out were imported into the DockThor online website for molecular docking. In addition, David database was used to perform gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the intersecting targets. Results The analysis showed that 110, 19, 154, 121, and 51 active components, respectively, were obtained for the above five classic prescriptions, and the numbers of drug targets were 7426, 1435, 9544, 6619, and 2427, respectively. Finally 4001 HCC disease targets were screened out. There were 260, 169, 276, 242, and 192 intersecting targets, respectively, between the five prescriptions and the HCC disease targets, and the survival analysis on the GEPIA online website obtained the common hub genes of PIK3CA, SRC, MAPK1, MAPK3 (all P < 0.05) and AKT1 (P > 0.05). Quercetin was the common active component of the five prescriptions, and isobavachin and Kanzonol W were the common active components of Xiaoyao powder combined with Sijunzi decoction, Longdan Xiegan decoction combined with Xiayuxue decoction, and Wupi decoction combined with Sijunzi decoction; the results of molecular docking showed that the above three components had a strong ability of binding to PIK3CA and SRC. GO enrichment analysis showed that these targets were involved in various biological processes including drug response, protein phosphorylation, inflammatory response, and angiogenesis, and KEGG enrichment analysis showed that the common pathways involved were cancer pathway, PI3K-Akt signaling pathway, MAPK signaling pathway, Ras signaling pathway, HIF-1 signaling pathway, hepatitis B pathway, and hepatitis C pathway. Conclusion Quercetin, isoflavone, and Kanzonol W have the potential mechanism of action involving multiple targets and pathways in the treatment of HCC.
- Carcinoma, Hepatocellular /
- Network Pharmacology /
- Signal Transduction

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图 1 5种经典方剂活性成分靶点与肝癌靶点映射图

注: a, 逍遥散合四君子汤; b, 茵陈蒿汤; c, 龙胆泻肝汤合下瘀血汤; d, 五皮饮合四君子汤; e, 一贯煎。

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图 2 5个靶蛋白基因表达与肝癌预后关系

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图 3 5种方剂关键活性成分与关键靶点分子对接评分统计图

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图 4 GO富集分析柱状图

注: a, 逍遥散合四君子汤; b, 茵陈蒿汤; c, 龙胆泻肝汤合下瘀血汤; d, 五皮饮合四君子汤; e, 一贯煎。

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图 5 KEGG通路富集分析气泡图

注: a, 逍遥散合四君子汤; b, 茵陈蒿汤; c, 龙胆泻肝汤合下瘀血汤; d, 五皮饮合四君子汤; e, 一贯煎。

下载: 全尺寸图片幻灯片

表 1 5种经典方剂度值靠前成分及其MOLID编号

项目	逍遥散合四君子汤	茵陈蒿汤	龙胆泻肝汤合下瘀血汤	五皮饮合四君子汤	一贯煎
自由度中位数	30	36	35	28	18
成分MOLID编号及度值	MOL004945 异补骨脂黄酮 (度值: 43)	MOL001406 藏花酸 (度值: 42)	MOL000552 Tenaxin I (度值: 44)	MOL004945 异补骨脂黄酮 (度值: 43)	MOL005384 苏奇内酯 (度值: 43)
	MOL004820 Kanzonol W (度值: 42)	MOL008041 Eupatolitin (度值: 41)	MOL002932 Panicolin (度值: 44)	MOL004820 Kanzonol W (度值: 42)	MOL000098 槲皮素 (度值: 38)
	MOL001689 刺槐素 (度值: 42)	MOL008040 泽兰素 (度值: 41)	MOL004945 异补骨脂黄酮 (度值: 43)	MOL000098 槲皮素 (度值: 41)	MOL002056 蛇菰宁 (度值: 37)
	MOL002881 香叶木素 (度值: 41)	MOL000354 异鼠李素 (度值: 41)	MOL002925 5, 7, 2′, 6′-四羟基黄酮 (度值: 43)		MOL009650 莨菪碱 (度值: 28)
	MOL000098 槲皮素 (度值: 38)	MOL000098 槲皮素 (度值: 38)	MOL004820 Kanzonol W (度值: 42)		MOL008647 N-反式-阿魏酸酪胺 (度值: 28)
			MOL000098 槲皮素 (度值: 41)

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表 2 5种经典方剂PPI网络中Hub基因度值

方剂	逍遥散合四君子汤	茵陈蒿汤	龙胆泻肝汤合下瘀血汤	五皮饮合四君子汤	一贯煎
靶点	PIK3CA (度值: 51)	SRC (度值: 36)	PIK3CA (度值: 49)	SRC (度值: 48)	MAPK1 (度值: 36)
	MAPK1 (度值: 48)	PIK3CA (度值: 36)	MAPK1 (度值: 48)	PIK3CA (度值: 48)	PIK3CA (度值: 35)
	SRC (度值: 48)	MAPK1 (度值: 34)	SRC (度值: 45)	MAPK1 (度值: 43)	AKT1 (度值: 34)
	AKT1 (度值: 43)	MAPK3 (度值: 28)	MAPK3 (度值: 44)	MAPK3 (度值: 42)	SRC (度值: 34)
	MAPK3 (度值: 43)	AKT1 (度值: 26)	AKT1 (度值: 43)	AKT1 (度值: 42)	MAPK3 (度值: 31)

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表 3 5种经典方剂关键活性成分与关键基因的分子对接结果

序号	活性成分	相对分子质量(g/mol)	基因(蛋白库编号)
序号	活性成分	相对分子质量(g/mol)	PIK3CA(3ZIM)	MAPK1(3W55)	MAPK3(4QTB)	SRC(3EL7)	LOXL2(5ZE3)
1	槲皮素	302.2	-7.384	-7.220	-6.522	-8.484	-6.460
2	异补骨脂黄酮	324.4	-7.398	-7.203	-7.109	-7.433	-7.219
3	Kanzonol W	336.3	-7.901	-7.454	-7.330	-9.191	-7.567

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