目的 探讨苗药马蹄金对对乙酰氨基酚（APAP）诱导的药物性肝损伤（DILI）的保护作用及分子机制。方法 采用网络药理学筛选马蹄金的主要成分及抗DILI潜在靶点，预测核心调控通路；通过分子对接验证关键成分与核心靶点的结合稳定性；将42只小鼠随机分为7组（n＝6）：对照组，模型组，N-乙酰半胱氨酸（NAC，阳性药，200 mg·kg^-1）组，马蹄金醇提物（DREE）低、中、高剂量（生药8.2、16.3、32.5 g·kg^-1）组，DREE单给药（仅给予32.5 g·kg^-1 DREE，不造模）组，ig给药7 d后，除对照组和DREE单给药组外，其余组ip APAP诱导DILI，24 h后取材。试剂盒法检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）水平和肝组织超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽（GSH）、丙二醛（MDA）水平；苏木精-伊红（HE）染色观察肝组织病理形态变化； Western blotting检测SRC/磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）通路相关蛋白（SRC、p-PI3K-p85、PI3K-p85、p-Akt、Akt）的表达。结果 网络药理学筛选出DREE的45个成分及163个抗DILI潜在靶点，PI3K-Akt通路为核心通路，Akt1、肿瘤坏死因子（TNF）、SRC为关键靶点；分子对接显示，DREE中β-谷甾醇、乌苏酸与SRC的结合能最低（分别为-55.1、-51.5 kJ·mol^-1），结合稳定性最优。动物实验显示，与模型组相比，DREE中、高剂量组血清ALT、AST水平显著降低（P＜0.05、0.01），肝组织SOD、CAT、GSH活性升高，MDA含量降低（P＜0.05、0.01），肝组织坏死及炎症浸润明显改善； Western blotting证实，DREE高剂量组可显著抑制肝组织SRC蛋白表达及PI3K、Akt的磷酸化水平（P＜0.05）。DREE单给药组各项指标与对照组比较无显著差异（P＞0.05），提示实验剂量DREE无明显肝毒性。结论 DREE对APAP诱导的DILI具有显著保护作用，其机制可能与抑制SRC/PI3K/Akt信号通路激活、缓解氧化应激损伤有关。

Objective To investigate the protective effect and molecular mechanism of Dichondra repens, a Miao ethnic medicine, against acetaminophen (APAP)-induced drug-induced liver injury (DILI). Methods Network pharmacology was used to screen the main components of D. repens and potential targets against drug-induced liver injury (DILI), and to predict the core regulatory signaling pathways. Molecular docking was performed to verify the binding stability between key components and core targets. An acetaminophen (APAP)-induced mouse model of DILI was established. A total of 42 mice were randomly divided into seven groups (n=6): control group, model group, N-acetylcysteine (NAC) (positive drug, 200 mg·kg^-1) group, D. repens ethanol extract (DREE) low, medium, and high dose (crude drug 8.2, 16.3, 32.5 g·kg^-1) groups, and DREE single administration group (only given 32.5 g·kg^-1 DREE without modeling). After 7 days of ig administration, except for the control group and DREE single administration group, the remaining groups were induced with DILI by ip APAP, and samples were taken after 24 h. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured, as well as the levels of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and malondialdehyde (MDA) in liver tissue. Pathomorphological changes in the liver were observed by hematoxylin-eosin (HE), and the expression of proteins related to the SRC/PI3K/Akt signaling pathway (SRC, p-PI3K-p85, PI3K-p85, p-Akt, Akt) was verified by Western blotting. Results Network pharmacology screening identified 45 components of DREE and 163 potential targets involved in its anti-DILI effect. The PI3K-Akt signaling pathway was identified as the core pathway, with Akt1, TNF, and SRC being the key targets. Molecular docking results showed that β-sitosterol and ursolic acid from DREE had the lowest binding energies with SRC (-55.1 and -51.5 kJ·mol^-1, respectively), indicating the optimal binding stability. Animal experiment results demonstrated that compared with the model group, the serum levels of ALT and AST in DREE medium and high dose groups were significantly decreased (P<0.05 and 0.01), the activities of SOD, CAT, and GSH in liver tissues were increased, and the MDA content was decreased (P<0.05 and 0.01). Meanwhile, hepatic necrosis and inflammatory infiltration were significantly improved in DREEtreated groups. Western blotting analysis confirmed that the high-dose DREE group could significantly inhibit the expression of SRC protein and the phosphorylation levels of PI3K and Akt in liver tissues (P<0.05). No significant differences were observed in all indicators between the DREE negative control group and the normal group (P > 0.05), suggesting that DREE at the experimental dose exerted no obvious hepatotoxicity. Conclusion D. repens exerts a significant protective effect against APAP-induced DILI. Its mechanism may be related to the inhibition of SRC/PI3K/Akt signaling pathway activation and the alleviation of oxidative stress injury.

R965

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（2024）13号］