Chen Aixin, Chen Tianhang, He Yu, Yang Jiehong, Wan Haitong
School of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China.
School of Basic Medical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Pathogens. 2025 Aug 21;14(8):834. doi: 10.3390/pathogens14080834.
This study aims to investigate the pharmacodynamic effects and underlying mechanisms of the Chinese herbal formula RuHaoDaShi (RHDS) granules against the influenza virus in experimental models.
This study aims to employ network pharmacology to identify the active components of RHDS and its potential targets and mechanisms of action against H1N1. The molecular docking approach validated the interactions between the core targets and the RHDS compounds. In vitro, the antiviral activity of RHDS was assessed by therapeutic, prophylactic, and premixed administration to H1N1-infected A549 cells. An in vivo experiment was conducted using a mouse H1N1 pneumonia model. The model was treated with a dose of 1.04, 2.08, and 4.16 g/kg of RHDS, administered via gavage daily. The study's objective was to evaluate the antiviral activity and mechanism of action of RHDS in mice. Mice were evaluated on day 6 by assessing survival, viral load (RT-qPCR), lung pathology (HE staining), inflammatory cytokines (ELISA, immunohistochemistry), and ferroptosis markers (WB, qPCR).
Network pharmacology identified 77 biologically active RHDS compounds (e.g., quercetin and kaempferol) and 32 core targets common to RHDS, H1N1, and ferroptosis. Molecular docking was used to verify a high affinity for binding between the core targets HIF-1α, MAPK3, and key RHDS compounds. In vitro studies demonstrated that RHDS exhibited protective properties against H1N1-infected cells, with the therapeutic delivery method proving the most efficacious. In vivo studies have shown that RHDS reduces mortality, lung index, and viral load in mice while attenuating histopathological damage. The study demonstrated a reduction in the release of inflammatory cytokines, including IL-6, IFN-γ, and IL-17A, and decreased expression levels of MPO and F4/80 proteins in lung tissue. Mechanistically, the administration of RHDS resulted in the up-regulation of the expression levels of GPX4, SLC7A11, and Nrf2 proteins while concomitantly inhibiting the expression of HIF-1α, COX2, and ACSL4. These findings confirm the modulatory effect of RHDS on the GPX4/SLC7A11/Nrf2 pathway.
RHDS demonstrated a protective effect against H1N1-induced cytopathy in vitro and was effective in attenuating H1N1-induced pneumonia in murine models. The study suggests that RHDS has antiviral potential to treat H1N1 viral pneumonia by modulating inflammatory cytokines and the GPX4/SLC7A11/Nrf2 pathway.
本研究旨在探讨中药复方乳蒿大实（RHDS）颗粒在实验模型中抗流感病毒的药效学作用及潜在机制。
本研究旨在运用网络药理学来鉴定RHDS的活性成分及其抗H1N1的潜在靶点和作用机制。分子对接方法验证了核心靶点与RHDS化合物之间的相互作用。在体外，通过对感染H1N1的A549细胞进行治疗性、预防性和预混给药来评估RHDS的抗病毒活性。使用小鼠H1N1肺炎模型进行体内实验。该模型每天通过灌胃给予1.04、2.08和4.16 g/kg剂量的RHDS。该研究的目的是评估RHDS在小鼠中的抗病毒活性和作用机制。在第6天通过评估存活率、病毒载量（RT-qPCR）、肺病理学（HE染色）、炎性细胞因子（ELISA、免疫组织化学）和铁死亡标志物（WB、qPCR）对小鼠进行评估。
网络药理学鉴定出77种具有生物活性的RHDS化合物（如槲皮素和山柰酚）以及RHDS、H1N1和铁死亡共有的32个核心靶点。分子对接用于验证核心靶点HIF-1α、MAPK3与关键RHDS化合物之间具有高亲和力的结合。体外研究表明，RHDS对感染H1N1的细胞具有保护作用，其中治疗性给药方法最为有效。体内研究表明，RHDS可降低小鼠的死亡率、肺指数和病毒载量，同时减轻组织病理学损伤。该研究表明炎性细胞因子（包括IL-6、IFN-γ和IL-17A）的释放减少，肺组织中MPO和F4/80蛋白的表达水平降低。从机制上讲，给予RHDS导致GPX4、SLC7A11和Nrf2蛋白的表达水平上调，同时抑制HIF-1α、COX2和ACSL4的表达。这些发现证实了RHDS对GPX4/SLC7A11/Nrf2通路的调节作用。
RHDS在体外对H1N1诱导的细胞病变具有保护作用，并且在减轻小鼠H1N1诱导的肺炎方面有效。该研究表明，RHDS具有通过调节炎性细胞因子和GPX4/SLC7A11/Nrf2通路治疗H1N1病毒性肺炎的抗病毒潜力。
