白术内酯Ⅲ通过调节巨噬细胞极化和可变聚腺苷酸化与非结构蛋白1结合以抑制甲型流感。

Atractylenolide-Ⅲ binds non-structural protein-1 to suppress influenza A by modulating macrophage polarization and alternative polyadenylation.

作者信息

Liang Jinlong, Guo Huiyi, Yang Yan, Liang Guorun, Chen Zhixuan, Li Dan, Liang Hao, Qiu Jiayin, Mei Qinghua, Liu Shuwen, Yang Jie, Wu Wenjiao

机构信息

Department of Pharmacy, The Affiliated Guangdong Second Provincial General Hospital of Jinan University, Guangzhou 510317, PR China; NMPA Key Laboratory for Research and Evaluation of Drug Metabolism, Guangdong Provincial Key Laboratory of New Drug Screening, Guangdong-Hongkong-Macao Joint Laboratory for New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, PR China.

NMPA Key Laboratory for Research and Evaluation of Drug Metabolism, Guangdong Provincial Key Laboratory of New Drug Screening, Guangdong-Hongkong-Macao Joint Laboratory for New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, PR China.

出版信息

Phytomedicine. 2025 Jun;141:156704. doi: 10.1016/j.phymed.2025.156704. Epub 2025 Mar 29.

DOI:10.1016/j.phymed.2025.156704

PMID:40220413

Abstract

BACKGROUND

Drug-resistant influenza demands novel antiviral treatments. Non-structural protein 1 (NS1) of influenza A virus (IAV) regulates the viral life cycle and host immune response, thus becoming a promising therapeutic target. The atractylenolide (ACT) -Ⅲ exhibits notable anti-IAV efficacy; however, its in vivo anti-IAV activity and the underlying mechanisms need further exploration.

PURPOSE

We explored the binding affinity of ACT-Ⅲ with NS1 and elucidated the in vivo anti-influenza activity and underlying mechanism of ACT-Ⅲ.

STUDY DESIGN AND METHODS

Surface plasmon resonance (SPR) analysis was utilized to determine the binding affinity of ACT-Ⅲ to NS1. The anti-IAV activity of ACT-Ⅲ was further tested in vitro using IAV-infected lung epithelial cells and in vivo in IAV-challenged mice. The evaluation criteria included the assessment of virus-associated cytopathies, viral protein and gene expression, virus titer, mice weight loss, survival curve, and lung pathology. Transcriptomic sequencing was conducted to evaluate the inflammation regulatory function of ACT-Ⅲ in IAV-infected Raw264.7 cells. Quantitative real-time PCR (qRT-PCR) was used to detect the target gene expression. Flow cytometry was utilized to assess macrophages polarization. Co-immunoprecipitation was employed to examine the impact of ACT-Ⅲ on the interaction between NS1 and cleavage and polyadenylation-specific factor 4 (CPSF4).

RESULTS

ACT-Ⅲ exhibited anti-IAV activity in vitro and in vivo, mitigating virus-induced lung injury. Mechanistically, ACT-Ⅲ impeded IAV infection and viral induced inflammation by binding to viral NS1, which reduced the activation of pro-inflammatory signaling, and disrupted the NS1-CPSF4 interaction that mediates alternative polyadenylation (APA).

CONCLUSION

ACT-Ⅲ inhibits IAV replication and viral pneumonia by binding to NS1, showing potential as a therapeutic agent for IAV treatment.

摘要

背景

耐药性流感需要新型抗病毒治疗方法。甲型流感病毒（IAV）的非结构蛋白1（NS1）调节病毒生命周期和宿主免疫反应，因此成为一个有前景的治疗靶点。白术内酯（ACT）-Ⅲ具有显著的抗IAV疗效；然而，其体内抗IAV活性及潜在机制有待进一步探索。

目的

我们探究了ACT-Ⅲ与NS1的结合亲和力，并阐明了ACT-Ⅲ的体内抗流感活性及潜在机制。

研究设计与方法

利用表面等离子体共振（SPR）分析确定ACT-Ⅲ与NS1的结合亲和力。使用IAV感染的肺上皮细胞在体外进一步测试ACT-Ⅲ的抗IAV活性，并在IAV攻击的小鼠体内进行测试。评估标准包括对病毒相关细胞病变、病毒蛋白和基因表达、病毒滴度、小鼠体重减轻、生存曲线和肺部病理学的评估。进行转录组测序以评估ACT-Ⅲ在IAV感染的Raw264.7细胞中的炎症调节功能。使用定量实时PCR（qRT-PCR）检测靶基因表达。利用流式细胞术评估巨噬细胞极化。采用免疫共沉淀法检测ACT-Ⅲ对NS1与切割及聚腺苷酸化特异性因子4（CPSF4）之间相互作用的影响。