| 引用本文:李芝奇，陈美琳，郭思敏，范琦琦，林子力，钟鑫悦，蔡琼，高睿涵，林瑞超，赵崇军，刘振权.基于斑马鱼模型结合网络药理学探究重楼肝毒性机制[J].世界中医药,2023,(06):. |
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| 基于斑马鱼模型结合网络药理学探究重楼肝毒性机制 |
| Hepatotoxicity Mechanism of Paridis Rhizoma Based on Zebrafish Model Combined with Network Pharmacology |
| 投稿时间：2021-12-16 |
| DOI：10.3969/j.issn.1673-7202.2023.06.001 |
| 中文关键词: 重楼 肝毒性 斑马鱼 网络药理学 分子对接 RT-PCR验证 PI3K-AKT信号通路 Ras信号通路 |
| English Keywords:Paridis Rhizoma Hepatotoxicity Zebrafish Network pharmacology Molecular docking RT-PCR PI3K-Akt signaling pathway Ras signaling pathway. |
| 基金项目:国家科技重大专项(2018ZX09735005)；北京中医药大学新教师启动基金项目(2020-JYB-XJSJJ-009)；北京中医药大学重点攻关项目(2020-JYB-ZDGG-035) |
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| 中文摘要: |
| 目的：重楼在《中华人民共和国药典》中记载为小毒，但其毒性机制尚未阐明。随着人们对中药安全性问题的重视，重楼的临床使用安全性及其机制也待明确，本文旨在阐明重楼提取物的肝毒性机制。方法：通过斑马鱼模式生物进行急性毒性验证，采用4 dpf斑马鱼暴露于重楼提取物中，24 h后吖啶橙染色，组织病理切片证明其肝脏毒性。通过网络药理学及分子对接技术预测重楼肝毒性靶点，并采用实时聚合酶链反应（RT-PCR）验证网络预测靶点及通路。结果：实验结果表明，重楼提取物暴露处理4 dpf斑马鱼24 h，吖啶橙染色重楼水提物组及醇提物组在肝脏部分均会出现黄绿色荧光点，组织病理切片显示肝细胞排列紊乱松散，出现细胞凋亡、空泡等现象，表明重楼具有肝脏毒性。网络药理学结果预测重楼肝毒性可能与细胞增殖的肉瘤基因(Ras)信号通路，能量代谢和信号转导相关的磷酸化磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、酪氨酸激酶（JAK）/信号转导与转录激活因子（STAT）信号通路相关，血管内皮生长因子A(VEGFA）、血小板衍生生长因子受体（PDGFRB）、鼠双微基因2（MDM2)、STAT3、雷帕霉素靶蛋白(MTOR）、原癌基因（MET)、B细胞淋巴瘤2样1（BCL2L1)、成纤维细胞生长因子2（FGF2）、白细胞介素-2（IL-2）及MAPK14可能是重楼造成肝毒性核心靶点，RT-PCR实验结果验证了以上推测。结论：重楼可能影响肝细胞增殖、能量代谢等通路的信号转导，从而引起肝毒性。 |
| English Summary: |
| Paridis Rhizoma is recorded as a minor poison in Chinese Pharmacopoeia，but its toxic mechanism has not been clarified.With the attention paid to the safety of Chinese medicine in recent years，the safety and mechanism of Paridis Rhizoma in clinical use also need to be clarified.The purpose of this paper is to clarify the hepatotoxicity mechanism of Paridis Rhizoma extract.Methods:The acute hepatotoxicity was verified by zebrafish.The zebrafish with 4 days of post-fertilization(4 dpf) were exposed to Paridis Rhizoma extract，acridine orange staining was carried out after 24 h，and the hepatotoxicity was proved by histopathological section.The hepatotoxicity targets of Paridis Rhizoma were predicted by network pharmacology and molecular docking technology，and real-time polymerase chain reaction(RT-PCR) was used to verify the network prediction targets and pathways.Results:According to the experimental results，yellow-green fluorescent spots appeared in the liver of zebrafish in the Paridis Rhizoma water extract group and alcohol extract group after exposure treatment for 24 h.Histopathological sections showed that the arrangement of hepatocytes was disordered and loose，and nuclear apoptosis and vacuoles appeared，indicating hepatotoxicity.The results of network pharmacology predicted that the hepatotoxicity of Paridis Rhizoma may be related to the Ras signaling pathway of cell proliferation，phosphoinositide 3-kinase/protein kinase B(PI3K/Akt) signaling pathway，mitogen-activated protein kinase(MAPK) signaling pathway，and Janus kinase-signal transducer and activator of transcription(JAK/STAT) signaling pathway related to energy metabolism and signal transduction.Vascular endothelial growth factor A(VEGFA)，platelet-derived growth factor receptor beta(PDGFRB)，murine double minute 2(MDM2)，signal transducers and activators of transcription 3(STAT3)，mammalian target of rapamycin(MTOR)，MET，Bcl-2-like protein 1(BCL2L1)，fibroblast growth factor 2(FGF2)，interleukin-2(IL-2)，and MAPK14 may be the core targets of hepatotoxicity caused by Paridis Rhizoma.The results of RT-PCR experiments verified the above speculation.Conclusion:Paridis Rhizoma may affect the signal transduction of hepatocyte proliferation and energy metabolism，which leads to hepatotoxicity. |
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