Pu XingNa, Lu Cheng, Yang Xing, He HongPing, Chen XingLong, Wang RuiRui, Li BaoJing, Chen Shuai, Zhang Yi, Wang WeiGuang, Li YanPing
Yunnan University of Chinese Medicine, Kunming, 650500, China.
Yunnan University of Chinese Medicine, Kunming, 650500, China.
Phytomedicine. 2025 Jan;136:156323. doi: 10.1016/j.phymed.2024.156323. Epub 2024 Dec 14.
The pathophysiology of liver diseases is significantly influenced by oxidative stress, making its alleviation a key strategy for treatment. The Keap1/Nrf2 signaling pathway is the body's most crucial antioxidant defense mechanism. Traditional Chinese medicine, Desmodium heterocarpon (L.) DC, has shown promising hepatoprotective effects, however, the specific active components and underlying mechanisms of its liver-protective properties remain inadequately understood. Further investigation into the bioactive constituents and mechanisms of its hepatoprotective action is therefore essential.
This study aims to identify the active ingredients in D. heterocarpon and to explore its hepatoprotective properties and underlying mechanisms.
The hepatoprotective activity of the ethyl acetate fraction (JEAE) from D. heterocarpon was first evaluated utilizing a mouse model of acute liver damage (ALI) caused by CCl. Molecular and histological analyses, including H&E staining, ELISA, and Western blot, were used to assess liver protection. The chemical constituents of JEAE were further identified using UPLC-MS/MS, and the molecular network of the JEAE fraction was analyzed. Compounds were isolated through column chromatography, and their antioxidant and hepatoprotective effects were assessed in an H₂O₂-induced HepG2 cell model using molecular assays. Additionally, binding interactions between active compounds and Keap1 were evaluated using molecular docking, molecular dynamics simulations, and surface plasmon resonance.
The ethyl acetate fraction of Desmodium heterocarpon (JEAE) showed remarkable antioxidant activity, with the highest flavonoid contents among extract fractions. In CCl₄-induced liver injury models, JEAE improved liver function, reduced ALT and AST levels, and enhanced antioxidant enzyme activities, suggesting hepatoprotective effects via the Keap1/Nrf2 pathway. 47 compounds were identified in JEAE, and fourteen flavonoids, including two novel compounds (1 and 2), were isolated from the JEAE fraction. Compounds 1, 3, 5, 8, and 14 notably protected HepG2 cells from oxidative damage, reduced ROS levels, and maintained mitochondrial function. These compounds also showed strong binding affinities to Keap1 and other antioxidant receptors, with molecular dynamics simulations confirming their stability and binding potential as effective hepatoprotective agents.
This study demonstrates that the ethyl acetate fraction of Desmodium heterocarpon (JEAE) exhibits significant hepatoprotective effects, largely attributed to its flavonoid-rich composition. The protective effects are mediated through antioxidant pathways, particularly the Keap1/Nrf2 signaling pathway. Newly identified isoflavanes and other flavonoids in JEAE show strong potential as bioactive compounds, with stability and binding affinities supporting their role in reducing oxidative stress. These findings suggest D. heterocarpon as a promising source of hepatoprotective agents and provide a foundation for further exploration of its therapeutic applications.
肝脏疾病的病理生理学受到氧化应激的显著影响，因此减轻氧化应激成为治疗的关键策略。Keap1/Nrf2信号通路是人体最关键的抗氧化防御机制。中药异叶山绿豆已显示出有前景的肝脏保护作用，然而，其肝脏保护特性的具体活性成分和潜在机制仍未得到充分了解。因此，进一步研究其生物活性成分及其肝脏保护作用的机制至关重要。
本研究旨在鉴定异叶山绿豆中的活性成分，并探索其肝脏保护特性及潜在机制。
首先利用四氯化碳诱导的急性肝损伤（ALI）小鼠模型评估异叶山绿豆乙酸乙酯部位（JEAE）的肝脏保护活性。采用苏木精-伊红染色、酶联免疫吸附测定（ELISA）和蛋白质印迹法等分子和组织学分析方法来评估肝脏保护作用。使用超高效液相色谱-串联质谱（UPLC-MS/MS）进一步鉴定JEAE的化学成分，并分析JEAE部位的分子网络。通过柱色谱分离化合物，并在过氧化氢诱导的HepG2细胞模型中使用分子分析方法评估其抗氧化和肝脏保护作用。此外，使用分子对接、分子动力学模拟和表面等离子体共振评估活性化合物与Keap1之间的结合相互作用。
异叶山绿豆乙酸乙酯部位（JEAE）表现出显著的抗氧化活性，在提取物部位中黄酮含量最高。在四氯化碳诱导的肝损伤模型中，JEAE改善了肝功能，降低了谷丙转氨酶（ALT）和谷草转氨酶（AST）水平，并增强了抗氧化酶活性，表明其通过Keap1/Nrf2途径发挥肝脏保护作用。在JEAE中鉴定出47种化合物，并从JEAE部位分离出14种黄酮类化合物，包括两种新化合物（化合物1和化合物2）。化合物1, 3, 5, 8和14显著保护HepG2细胞免受氧化损伤，降低活性氧（ROS）水平，并维持线粒体功能。这些化合物还显示出与Keap1和其他抗氧化受体的强结合亲和力，分子动力学模拟证实了它们作为有效肝脏保护剂的稳定性和结合潜力。
本研究表明，异叶山绿豆乙酸乙酯部位（JEAE）具有显著的肝脏保护作用，这主要归因于其富含黄酮类化合物的组成。保护作用是通过抗氧化途径介导的，特别是Keap1/Nrf2信号通路。JEAE中新鉴定的异黄酮和其他黄酮类化合物显示出作为生物活性化合物的强大潜力，其稳定性和结合亲和力支持它们在减轻氧化应激中的作用。这些发现表明异叶山绿豆是一种有前景的肝脏保护剂来源，并为进一步探索其治疗应用提供了基础。
