目的 开展博落回醇提物（MCEE）对斑马鱼和HepG2细胞的肝毒性评价，通过网络药理学和分子对接预测主要活性成分血根碱和白屈菜红碱肝毒性潜在机制。方法 考察MCEE (5、10、20、40、80、160、320、640μg·mL^-1)对受精4 d斑马鱼幼鱼死亡率的影响，观察肝脏病理组织病变，测定丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）活性；采用MTT、Hoechst 33342染色法评价MCEE (5、10、20、40、80、160、320、640μg·mL^-1)对HepG2细胞增殖和细胞凋亡作用。通过Swiss Target Prediction数据库、Gene Cards、OMIM和MalaCards数据库预测白屈菜红碱、血根碱和肝毒性交集靶点，STRING数据库构建蛋白质-蛋白质互相作用（PPI），得到核心靶基因；通过David数据库进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析，利用Autodock Vina软件对核心靶点与白屈菜红碱、血根碱进行分子对接验证。结果 MCEE的10%致死质量浓度(LC₁₀)为6.07μg·mL^-1；与对照组相比，MCEE可使斑马鱼肝脏受损，肝脏组织中出现大量空泡，肝细胞排列紊乱、松散，甚至出现组织结构降解，AST和ALT活性均显著升高（P<0.01）；与对照组相比，MCEE抑制HepG2细胞增殖（P<0.05、0.01），诱导细胞凋亡。网络药理学结果表明血根碱、白屈菜红碱和肝毒性交集靶点有15个，PPI网络核心靶点为GSK3B、MAPK1、MTOR、PIK3CG、PTGS2等；GO功能富集靶点主要富集于TORC2信号负调控细胞对营养的反应、线粒体外膜、MAP激酶活性等过程；KEGG通路富集分析发现血根碱、白屈菜红碱肝毒性与VEGF信号通路、m TOR信号通路、PI3K-Akt信号通路等有关。分子对接结果显示白屈菜红碱、血根碱与关键靶蛋白GSK3B、MAPK1、MTOR、PIK3CG、PTGS2之间有良好的结合作用。结论 MCEE具有一定的肝毒性，可能与TORC2信号负调控细胞对营养的反应、线粒体外膜、MAP激酶活性等过程，VEGF信号通路、m TOR信号通路、PI3K-Akt信号通路等有关。

Objective The hepatotoxicity of the methanol extract of Macleaya cordata（MCEE） on zebrafish and HepG2 cells was evaluated. The potential mechanisms of the hepatotoxicity of the main active components, sanguinarine and chelerythrine, were predicted through network pharmacology and molecular docking. Methods The effects of MCEE(5, 10, 20, 40, 80, 160, 320, 640 μg·mL^-1) on the mortality of 4-day-old zebrafish larvae were investigated, and the liver pathological tissue lesions were observed. The activities of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） were determined. The effects of MCEE(5, 10, 20, 40, 80, 160, 320, 640 μg·mL^-1) on the proliferation and apoptosis of HepG2 cells were evaluated by MTT and Hoechst 33342 staining. The intersection targets of sanguinarine and chelerythrine hepatotoxicity were predicted through the Swiss Target Prediction database, Gene Cards, OMIM and MalaCards databases. The protein-protein interaction network（PPI） was constructed by the STRING database to obtain the core target genes. The Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment analyses were performed through the David database. The molecular docking was verified by the Autodock Vina software between the core targets and MCEE. Results The 10% lethal concentration(LC₁₀) of MCEE was 6.07 μg·mL^-1. Compared with the control group, MCEE could cause liver damage in zebrafish, with a large number of vacuoles in the liver tissue, disordered and loose arrangement of liver cells, and even degradation of tissue structure. The activities of AST and ALT were significantly increased（P ＜ 0.01）. Compared with the control group, MCEE inhibited the proliferation of HepG2 cells（P ＜ 0.05, 0.01） and induced apoptosis. The network pharmacology results showed that there were 15 intersection targets of sanguinarine, chelerythrine and hepatotoxicity. The core targets of the PPI network were GSK3B, MAPK1, MTOR, PIK3 CG, PTGS2, etc. The GO functional enrichment targets were mainly enriched in the processes of negative regulation of TORC2 signaling in response to nutrients, mitochondrial outer membrane, MAP kinase activity, etc. The KEGG pathway enrichment analysis found that the hepatotoxicity of sanguinarine and chelerythrine was related to the VEGF signaling pathway, mTOR signaling pathway, PI3K-Akt signaling pathway, etc. The molecular docking results showed that sanguinarine and chelerythrine had good binding effects with the key target proteins GSK3B, MAPK1, MTOR, PIK3 CG, PTGS2. Conclusion MCEE has certain hepatotoxicity, which may be related to the processes of negative regulation of TORC2 signaling in response to nutrients, mitochondrial outer membrane, MAP kinase activity, etc., as well as the VEGF signaling pathway, mTOR signaling pathway, PI3K-Akt signaling pathway, etc.

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贵州省科技厅基础研究计划一般项目（黔科合基础-ZK[2022]一般472）；贵州中医药大学大学生创新创业训练计划项目（贵中医大创合字［2022］ 78号）；地方病与少数民族性疾病教育部重点实验室（贵州医科大学）开放课题基金资助项目（黔教合KY字［2019］047号）；贵州省高等学校中药民族药（苗药）新剂型新制剂工程研究中心项目（黔教技［2022］ 022号）