Key Laboratory of Forest Plant Ecology, Ministry of Education, Northeast Forestry University, Harbin 150040, China; Engineering Research Center of Forest Bio-Preparation, Ministry of Education, Northeast Forestry University, Harbin 150040, PR China; College of Chemistry, Chemical Engineering and Resource Utilization, Northeast Forestry University, Harbin 150040, China.
Department of Pharmaceutical Biology, Institute of Pharmaceutical and Biomedical Sciences, University of Mainz, 55128, Mainz, Germany.
Phytomedicine. 2022 Jul 20;102:154170. doi: 10.1016/j.phymed.2022.154170. Epub 2022 May 14.
Hepatic inflammation can substantially impact the development of acute hepatitis. It is a pressing need to identify and exploit novel therapeutic targets as well as effective drug therapies against acute hepatitis. Aucubin (AU) is one of the main active components extracted from the leaves of Eucommia ulmoides and possesses significant anti-inflammatory and antioxidant activities. However, the protective effect and mechanism of AU on acute hepatitis have not been reported yet.
This study aims to investigate the protective effect of AU on LPS-induced acute hepatitis and the mechanism of action.
The limma package was used to analyze differentially expressed genes (DEGs) between LPS-induced acute hepatitis and normal groups based on Gene Expression Omnibus (GEO) microarray data. Network pharmacology predicted targets for AU therapy against acute hepatitis, and Gene Ontology (GO) enrichment analysis of the biological processes involved in these targets. The key pathways were analyzed by protein-protein interaction, KEGG (Kyoto Encyclopedia of Genes and Genomes), and GSEA (Gene Set Enrichment Analysis) enrichment. The important interaction targets between AU and key pathways were evaluated by molecular simulation. The in silico predicted mechanism was verified based on in vitro and in vivo experiments.
A total of 116 intersection targets between AU prediction targets and differentially expressed genes were identified. They were functionally involved in the imbalance of "inflammation-anti-inflammation" and "oxidation-antioxidation" systems in the process of LPS-induced cases. In vitro experiments revealed that AU reduced inflammation in LPS-induced HepG2 cells by reducing the inflammatory cytokines TNF-α, IL-6, as well as iNOS enzyme activity levels. In addition, LPS-induced oxidative stress can be alleviated by AU via adjusting the levels of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Malone dialdehyde (MDA) and reactive oxygen species (ROS). Protein-protein interaction and GSEA results showed that AU might exert anti-inflammatory effects mainly through the STAT3/NF-κB signal pathway. Molecular dynamics simulation as well as in vivo tests further demonstrated AU restrained nuclear transfer of NF-κB (P65), probably through reducing phosphorylation of STAT3. In addition, AU appears to reduce oxidative stress by upregulating NRF2/HO-1.
We explored potential targets and signal pathways of AU in inhibiting acute hepatitis. AU exerted anti-inflammatory and antioxidant activities and may be a useful candidate drug for the treatment of acute hepatitis.
肝脏炎症会显著影响急性肝炎的发展。因此，有必要寻找和利用新型治疗靶点，并开发针对急性肝炎的有效药物疗法。梓醇（AU）是从杜仲叶中提取的主要活性成分之一，具有显著的抗炎和抗氧化活性。然而，AU 对急性肝炎的保护作用及其机制尚未见报道。
本研究旨在探讨 AU 对脂多糖（LPS）诱导的急性肝炎的保护作用及其作用机制。
基于基因表达综合数据库（GEO）微阵列数据，使用 limma 包分析 LPS 诱导的急性肝炎组和正常组之间的差异表达基因（DEGs）。通过网络药理学预测 AU 治疗急性肝炎的作用靶点，并对这些靶点涉及的生物学过程进行基因本体论（GO）富集分析。通过蛋白质-蛋白质相互作用、京都基因与基因组百科全书（KEGG）和基因集富集分析（GSEA）富集分析来分析关键途径。通过分子模拟评估 AU 与关键途径之间的重要相互作用靶点。基于体外和体内实验验证基于计算的预测机制。
共鉴定出 AU 预测靶点和差异表达基因之间的 116 个交集靶点。它们在 LPS 诱导的情况下，参与了“炎症-抗炎”和“氧化-抗氧化”系统失衡的过程。体外实验表明，AU 通过降低炎症细胞因子 TNF-α、IL-6 和诱导型一氧化氮合酶（iNOS）酶活性水平，减轻 LPS 诱导的 HepG2 细胞炎症。此外，AU 通过调节超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、丙二醛（MDA）和活性氧（ROS）水平，减轻 LPS 诱导的氧化应激。蛋白质-蛋白质相互作用和 GSEA 结果表明，AU 可能主要通过 STAT3/NF-κB 信号通路发挥抗炎作用。分子动力学模拟和体内试验进一步证明，AU 通过抑制 NF-κB（P65）的核转位来抑制炎症，这可能是通过降低 STAT3 的磷酸化来实现的。此外，AU 可能通过上调 NRF2/HO-1 来减轻氧化应激。
我们探讨了 AU 抑制急性肝炎的潜在靶点和信号通路。AU 发挥抗炎和抗氧化作用，可能是治疗急性肝炎的一种有用的候选药物。
