Co-construction Collaborative Innovation Center for Chinese Medicine Resources Industrialization by Shaanxi & Education Ministry, State Key Laboratory of Research, & Development of Characteristic Qin Medicine Resources (Cultivation), Shaanxi Innovative Drug Research Center, Shaanxi University of Chinese Medicine, Xianyang, Shaanxi 712083, PR China.
College of Pharmacy, Shaanxi University of Chinese Medicine, Xi'an, Shaanxi 712046, PR China.
Phytomedicine. 2024 Oct;133:155910. doi: 10.1016/j.phymed.2024.155910. Epub 2024 Jul 22.
Total flavonoids from Astragali Complanati Semen (TFACS), the main active ingredients in Astragali Complanati Semen (ACS), have been shown to have a protective effect on chronic liver injury (CLI), but the hepatoprotective targets and signalling pathways involved are unclear.
The aim of our study was to identify the anti-CLI targets and signalling pathways of TFACS and to comprehensively elucidate its mechanism of action via proteomics analysis combined with in vivo and in vitro experiments.
A CLI mouse model was generated via intraperitoneal injection of carbon tetrachloride (CCl) (CCl: olive oil = 1:4, 2 ml/kg, twice a week for 6 weeks). The hepatoprotective effect of TFACS was assessed by observing the pathological structure of the liver and analysing indicators of liver function. The key pathways and targets related to the hepatoprotective effect of TFACS were identified via 4D-label-free quantitative proteomics technology and further verified via in vivo indicator validation and in vitro cell experiments.
TFACS administration significantly normalized the histopathological structure and function of the liver, decreased the levels of inflammatory factors and oxidative stress indicators, and reduced the iron staining area and the levels of hepcidin and iron in the liver compared with those in the CLI model. A total of 424 differentially expressed proteins (DEPs) were identified between the TFACS and model groups, and these DEPs were enriched in the focal adhesion, PI3K-Akt, and ferroptosis pathways. Akt1, Pik3ca, NF-κB p65, Itga5, Itgb5, Itga6, Prkca, Fn1, Tfrc, and Vdac3 were identified as key targets of TFACS. TFACS administration significantly reversed the changes in the gene and protein expression of the key targets compared with those in the model group. In addition, TFACS treatment significantly reduced the levels of inflammatory cytokines and inhibited Akt1, NF-κB p65 and FAK activation in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. In an erastin-induced l-O2 ferroptosis cell model, treatment with TFACS normalized the mitochondrial structure, reduced the protein levels of Tfrc and Vdac3, inhibited lipid peroxidation, and reduced the amount of Fe in the mitochondria.
TFACS protected against CLI, and its mechanism of action may be related to inhibition of the focal adhesion, PI3K/Akt and ferroptosis signalling pathways.
来自黄芪种子总黄酮（TFACS）的主要活性成分黄芪种子（ACS）已被证明对慢性肝损伤（CLI）具有保护作用，但涉及的肝保护靶标和信号通路尚不清楚。
本研究旨在通过蛋白质组学分析结合体内和体外实验，鉴定 TFACS 的抗 CLI 靶标和信号通路，并全面阐明其作用机制。
采用腹腔注射四氯化碳（CCl）（CCl：橄榄油=1：4，2 ml/kg，每周两次，共 6 周）建立 CLI 小鼠模型。通过观察肝脏的病理结构和分析肝功能指标来评估 TFACS 的肝保护作用。通过 4D-无标记定量蛋白质组学技术鉴定与 TFACS 肝保护作用相关的关键途径和靶标，并通过体内指标验证和体外细胞实验进一步验证。
TFACS 给药可显著使肝的组织病理学结构和功能正常化，降低炎症因子和氧化应激指标的水平，并降低肝脏铁染色面积以及铁调素和铁的水平，与 CLI 模型相比。TFACS 组和模型组之间共鉴定出 424 个差异表达蛋白（DEPs），这些 DEPs 富集在粘着斑、PI3K-Akt 和铁死亡途径中。Akt1、Pik3ca、NF-κB p65、Itga5、Itgb5、Itga6、Prkca、Fn1、Tfrc 和 Vdac3 被鉴定为 TFACS 的关键靶标。与模型组相比，TFACS 给药可显著逆转关键靶标基因和蛋白表达的变化。此外，TFACS 治疗可显著降低脂多糖（LPS）诱导的 RAW 264.7 巨噬细胞中炎症细胞因子的水平，并抑制 Akt1、NF-κB p65 和 FAK 的激活。在依马替尼诱导的 l-O2 铁死亡细胞模型中，TFACS 处理可使线粒体结构正常化，降低 Tfrc 和 Vdac3 蛋白水平，抑制脂质过氧化，减少线粒体中的铁含量。
TFACS 可预防 CLI，其作用机制可能与抑制粘着斑、PI3K/Akt 和铁死亡信号通路有关。
