基于网络药理学的蒙药肋柱花抗急性肝损伤作用机制研究

doi:10.5246/jcps.2023.04.024

中国药学(英文版) ›› 2023, Vol. 32 ›› Issue (4): 268-282.DOI: 10.5246/jcps.2023.04.024

基于网络药理学的蒙药肋柱花抗急性肝损伤作用机制研究

敖民¹, 包明兰¹^,⁴^,^*(), 侯亚星¹, 月英¹, 李慧芳¹, 吴国华², 苏日嘎拉图³

1. 内蒙古民族大学蒙医药学院, 内蒙古通辽 028000
2. 内蒙古民族大学附属医院, 内蒙古通辽 028000
3. 赤峰学院, 内蒙古赤峰 024000
4. 蒙医药研发工程教育部重点实验室, 内蒙古通辽 028000

收稿日期:2022-09-12 修回日期:2022-11-17 接受日期:2023-01-29 出版日期:2023-04-29 发布日期:2023-04-29
通讯作者: 包明兰
作者简介:
+ Tel.: +86-13624757544, E-mail: baominglan@126.com
基金资助:
National Natural Science Foundation of China (Grant No. 81603386); Montr&D subject of the open fund, Engineering Laboratory of Inner Mongolia Autonomous Region (Grant No. MDK2018064); 2019 Double First in Class Special Project of Inner Mongolia Autonomous Region-innovative Team Construction of Mongolian Medicine Processing (Grant No. MY20190004).

Study on the mechanism of Mongolian medicine Herba Lomatognii against acute liver injury based on network pharmacology

Min Ao¹, Minglan Bao¹^,⁴^,^*(), Yaxing Hou¹, Ying Yue¹, Huifang Li¹, Guohua Wu², Su Ri Ga La Tu³

1 School of Mongolian Medicine, Inner Mongolia Minzu University, Tong Liao 028000, Inner Mongolia, China
2 Affiliated Hospital of Inner Mongolian Minzu University, Tong Liao 028000, Inner Mongolia, China
3 Chifeng university, Chifeng 024000, Inner Mongolia, China
4 Key Laboratory of Mongolian medicine research and development engineering, Ministry of Education. Tong Liao 028000, Inner Mongolia, China

Received:2022-09-12 Revised:2022-11-17 Accepted:2023-01-29 Online:2023-04-29 Published:2023-04-29
Contact: Minglan Bao

摘要/Abstract

摘要：

本文通过网络药理学和生物信息学研究蒙药肋柱花抗急性肝损伤作用机制, 为进一步研究肋柱花治疗急性肝损伤等代谢性疾病提供参考。应用肋柱花抗D-GLa诱导的急性肝损伤模型大鼠, 分析血清转氨酶和酶联免疫吸附法变化。采用TCMSP数据库及文献检索收集肋柱花化学成分及其对应的靶点, 应用GeneCards和DisGeNET疾病数据库获取急性肝损伤(ALI)靶点, 将肋柱花活性成分靶点与ALI靶点交集, 所得到的靶点为肋柱花抗急性肝损伤ALI的靶点。利用Metascape数据库对共有靶点进行GO功能及KEGG通路富集分析, 使用Cytoscape软件构建相关网络图, 筛选主要核心成分及关键靶点进行分子对接验证。并用蛋白免疫印迹法(Western Blot)进一步检测大鼠肝脏中转录激活因子3(STAT3)、AKT1、CASP3的表达情况。血清转氨酶和酶联免疫结果显示肋柱花对D-GLaN诱导的大鼠肝损伤有不同程度的保护作用和抗炎作用。网络药理学筛选共得到肋柱花的10个活性成分及289个对应的靶点, 急性肝损伤靶点843个, 二者共同靶点89个, GO富集和KEGG Pathway分析发现共同靶点主要为癌症通路、PI3K-Akt信号通路、Ras信号通路、HIF-1信号通路、TNF-α信号通路、Toll样受体信号通路、MAPK信号通路、VEGF信号通路、p53信号通路、NF-κB信号通路等。分子对接结果显示, 獐牙菜苦苷、木犀草素、山奈酚等核心成分能与EGFR、SRC、AKT1、STAT3、CASP3等靶点良好对接。蛋白免疫印迹结果显示, 与正常对照组对比, 模型组大鼠肝组织中的AKT1、STAT3和CASP3蛋白表达明显增加, 具有统计学意义(P < 0.05)。与模型组对比, 阳性组和肋柱花低、中、高剂量组肝组织中的AKT1、STAT3蛋白表达降低, 具有统计学意义(P < 0.05)。与模型组对比, 肋柱花中、高剂量组肝组织中的CASP3蛋白表达明显降低, 肋柱花低剂量组肝组织中CASP3蛋白表达未见明显差异(P > 0.05)。肋柱花是潜在治疗ALI的蒙药, 通过多靶点、多途径干涉ALI的进展, 其发挥治疗作用的机制可能与干预(STAT3)、AKT1、CASP3等靶点蛋白的表达及调控癌症信号通路、PI3K-Akt信号通路、TNF-α等信号通路进而抑制急性肝损伤和炎症反应有关。

关键词: 肋柱花, 急性肝损伤, 网络药理学, 分子对接, 蛋白免疫印迹

Abstract:

In the present study, we aimed to study the action mechanism of Mongolian medicine Herba Lomatognii on acute liver injury (ALI) using network pharmacology and bioinformatics methods to provide a reference for further study of Herba Lomatognii on metabolic diseases. Herba Lomatognii showed protective effects on D-GlaN-induced ALI model rats, which were used to analyze serum transaminase and enzyme-linked immunosorbent assay changes. The chemical constituents of Herba Lomatognii and their corresponding targets were collected by TCMSP database and literature search. ALI targets were compiled by GenCards and DisGeNet disease databases, and the targets of active constituents of Herba Lomatognii were interesting ALI targets. The obtained targets were targets of Herba Lomatognii against ALI. The Metascape database was used to do a Go function and KEGG pathway enrichment analysis of consensus targets, and Cytoscape software was used to create a correlation network diagram. Validation of molecular docking was carried out by analyzing important core components and significant targets. In addition, Western blotting analysis was utilized to detect transcription activator 3 (STAT3), AKT1, and CASP3 expressions in the rat liver. Serum transaminases and enzyme-linked immunostaining assay demonstrated that Herba Lomatognii had preventive and anti-inflammatory effects on D-GlaN-induced ALI in rats. From network pharmacology screening, a total of 10 active ingredients of Herba Lomatognii and 289 corresponding targets were identified, 843 targets of ALI and 89 targets of both, and GO enrichment and KEGG pathway analysis revealed that the common targets were mostly related to cancer, PI3K-Akt signaling pathway, Ras signaling pathway, HIF-1 signaling pathway, TNF-α signaling pathway, toll-like receptor signaling pathway, MAPK signaling pathway, VEGF signaling pathway, p53 signaling pathway, and NF-κB signaling pathway. Molecular docking results showed that the core components, such as swetiamain, luteolin, and kaempferol, could dock well with the targets of EGFR, SRC, AKT1, STAT3, and CASP3. The expressions of AKT1, STAT3, and CASP3 proteins in the liver tissues of model rats were considerably higher compared with the normal control rats based on Western blotting data (P < 0.05). In contrast to the model group, the AKT1 and STAT3 protein expressions in the liver tissues of the positive group and the Herba Lomatognii low-, medium-, and high-dose groups were significantly decreased (P < 0.05). Compared with the model group, the CASP3 protein expression in the liver tissue of the middle- and high-dosage groups of Herba Lomatognii was significantly lower, and no significant difference was observed in the liver tissue of the low-dosage group of Herba Lomatognii (P > 0.05). Herba Lomatognii contained potential therapeutic agents for ALI and could interfere with the development of ALI through multi-target and multi-pathway approaches.

Key words: Herb Lomatognii, Acute liver injury, Network pharmacology, Molecular docking, Western blotting

Supporting:

敖民, 包明兰, 侯亚星, 月英, 李慧芳, 吴国华, 苏日嘎拉图. 基于网络药理学的蒙药肋柱花抗急性肝损伤作用机制研究[J]. 中国药学(英文版), 2023, 32(4): 268-282.

Min Ao, Minglan Bao, Yaxing Hou, Ying Yue, Huifang Li, Guohua Wu, Su Ri Ga La Tu. Study on the mechanism of Mongolian medicine Herba Lomatognii against acute liver injury based on network pharmacology[J]. Journal of Chinese Pharmaceutical Sciences, 2023, 32(4): 268-282.

图/表 13

Table 1. Effects of Herba Lomatognii on serum transaminases (mean ± SEM, n = 10).

Figure 1. Effects of Herba Lomatognii on the contents of IL-6, IL-10, and TNF-α. Note: ##P < 0.01 vs. Control, **P < 0.01 vs. Model.

Figure 2. Veen diagram.

Figure 3. Network of potential targets of Herb Lomatognii treatment for ALI analyzed by STRING.

Figure 4. The core targets.

Table 2. Putative targets of Herba Lomatogni against ALI.

Table 3. Topological parameters of targets.

Figure 5. GO and KEGG enrichment analysis. (A) GO enrichment of Herba Lomatognii; (B) KEGG pathway enrichment.

Figure 6. Target-pathway network.

Table 4. Molecular docking results.

Figure 7. 3D diagrams of molecular models of Herba Lomatognii with predicted targets of AKT1 (A), STAT3 (B), EGFR (C), CASP3 (D), and SRC (E).

Table 5. Expressions of AKT1, STAT3, and CASP3 upon treatment of Herba Lomatognii (mean ± SEM, n = 10).

Figure 8. Electrophoresis of AKT1, STAT3, and CASP3 proteins in rat liver tissue of Herba Lomatognii groups.

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基于网络药理学的蒙药肋柱花抗急性肝损伤作用机制研究

Study on the mechanism of Mongolian medicine Herba Lomatognii against acute liver injury based on network pharmacology

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