Prunin inhibits Senecavirus A replication through the internal ribosome entry site in vivo and in vitro

doi:10.1016/j.jia.2026.05.018

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Prunin inhibits Senecavirus A replication through the internal ribosome entry site in vivo and in vitro

Han Xu¹, Longbin He¹, Xiaoxuan Nie¹, Jian Sun^{1, 2}, Chao Liu¹, Qinyue Lu¹, Anyu Li¹, Yongtao Zhao¹, Justin Jang Hann Chu³, Hongbo Cao⁴, Limin Li¹, Kuan Zhao¹, Wanzhe Yuan¹, Jianke Wang¹^#

¹Hebei Veterinary Biotechnology Innovation Center, College of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, China

²Weihai Marine Organism Health Promotion Center, Weihai Ocean Vocational College, Rongcheng 264300, China

³Department of Microbiology and Immunology and Infectious Disease Translation Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.

⁴College of Horticulture, Hebei Agricultural University, Baoding, Hebei 071000, China.

Highlights

l Prunin inhibits Senecavirus A (SVA) replication in a dose-dependent manner in vitro.

l For the first time, hnRNP A2B1, hnRNP K, and SAM68 are identified as ITAFs that bind to the SVA internal ribosome entry site (IRES).

l Prunin disrupts the interaction of these ITAFs with the SVA IRES, thereby suppressing viral translation.

Abstract
References

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摘要

A型塞内卡病毒（Senecavirus A，SVA）是一种新发猪源性病原体，可引起猪只水疱性病变、腹泻和死亡，给全球养猪业造成重大经济损失。目前尚无特异性抗病毒药物，开发新型抗病毒防控策略迫在眉睫。普鲁宁是一种天然黄酮苷类化合物，前期研究发现其可抑制内部核糖体进入位点（IRES）并具有抗病毒活性，但其对SVA的作用尚未见报道。本研究旨在系统评价普鲁宁对SVA的体内外抑制作用，并深入解析其是否通过靶向IRES介导的翻译过程发挥抗病毒效应。

体外细胞毒性试验结果显示，普鲁宁在BHK-21细胞中表现出较低的毒性作用。抗病毒活性检测表明，普鲁宁可剂量依赖性抑制SVA的复制。为明确普鲁宁在病毒复制周期中的作用环节，评估了普鲁宁对病毒生命周期各阶段的影响，结果表明普鲁宁主要靶向病毒复制的胞内阶段，能够显著下调病毒RNA的转录水平，而对病毒的吸附、直接灭活及子代病毒释放等阶段的病毒RNA水平无明显干扰作用，提示其可能作用于病毒进入细胞后的胞内复制关键环节。

Western blot和间接免疫荧光试验证实，普鲁宁可剂量依赖性抑制病毒结构蛋白的表达，感染细胞的比例亦相应减少。为阐明其分子作用靶点，利用IRES依赖性双荧光素酶报告系统研究发现，普鲁宁特异性靶向SVA的IRES翻译起始功能。RNA pull-down试验进一步证实，普鲁宁可干扰病毒IRES与宿主关键反式作用因子（IRES-transacting factors, ITAFs）之间的相互作用，阻断IRES-ITAF复合物组装，从而在翻译起始阶段抑制病毒蛋白合成。

体内试验进一步验证了普鲁宁的抗病毒效果。通过建立BALB/c小鼠感染模型，分别设计预防性给药和治疗性给药方案，结果表明普鲁宁均能显著降低SVA感染后小鼠肝、脾、肾及小肠中的病毒载量，Western blot检测证实各组织中病毒蛋白表达水平显著下降。与病毒对照组相比，普鲁宁处理组小鼠的病理损伤呈剂量依赖性地减轻，体重下降趋势有效减缓；药物对照组未见明显毒性反应，表明普鲁宁在有效剂量范围内安全性良好。

综上所述，普鲁宁通过特异性靶向IRES介导的翻译过程这一病毒蛋白合成的关键环节，有效抑制SVA复制，并在体内外均表现出良好的抗病毒效果。本研究发现普鲁宁作为新型IRES抑制剂具有抗病毒潜力，揭示其通过干扰IRES-ITAF相互作用发挥抗病毒效应的新机制，为后续开发针对SVA及其他IRES依赖性病毒的抗病毒药物奠定了重要理论基础。

Abstract

Senecavirus A (SVA) is an emerging swine pathogen that causes vesicular lesions, diarrhea, and mortality, resulting in substantial economic losses worldwide. Currently, no commercial vaccines or specific antiviral treatments are available, underscoring the urgent need for innovative strategies to control SVA. Prunin, a natural compound previously identified as an inhibitor of the internal ribosomal entry site (IRES) with antiviral activity, remains unexplored in SVA. Herein, we investigated the anti-SVA activity of prunin and explored its underlying mechanism. In vitro, prunin exhibited low cytotoxicity in BHK-21 cells and potently suppressed SVA replication in a dose-dependent manner. Time-of-addition experiments revealed that prunin primarily targeted the replication phase, with negligible effects on viral adsorption or release. Mechanistic investigations using an IRES-dependent dual-luciferase reporter system demonstrated that prunin significantly impaired IRES-driven translation. Furthermore, RNA pull-down assays indicated that prunin disrupted the interaction between the SVA IRES and key trans-acting factors, including hnRNP A2B1, hnRNP K, and SAM68. In vivo, both prophylactic and therapeutic administration of prunin in BALB/c mice markedly reduced viral loads compared to control groups. Together, these results indicate that prunin inhibits SVA replication by specifically targeting IRES-mediated translation-a critical step in viral protein synthesis. Our study identifies prunin as a novel IRES-targeting antiviral candidate and provides a strategic foundation for developing therapies against SVA.

Keywords: SVA prunin IRES antiviral mechanism viral replication translation inhibition

Online: 22 May 2026

Fund:

This work was supported by the Yanzhao Huangjintai Talent Gathering Plan Fund of Hebei Province (B2025010), National Natural Science Foundation of China (32373001) and Hebei Provincial Department of Agriculture and Rural Affairs (HBCT2024160204). We are grateful to members of the Wang laboratory for technical support and valuable discussions.

About author: Han Xu, E-mail: xh1940783074@163.com; #Correspondence Jianke Wang, E-mail: wangjianke@hebau.edu.cn

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Han Xu, Longbin He, Xiaoxuan Nie, Jian Sun, Chao Liu, Qinyue Lu, Anyu Li, Yongtao Zhao, Justin Jang Hann Chu, Hongbo Cao, Limin Li, Kuan Zhao, Wanzhe Yuan, Jianke Wang. 2026. Prunin inhibits Senecavirus A replication through the internal ribosome entry site in vivo and in vitro. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.05.018

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[3]	MA Shou-chen, LI Feng-min, YANG Shen-jiao, LI Chun-xi, XU Bing-cheng , ZHANG Xu-cheng. Effects of Root Pruning on Non-Hydraulic Root-Sourced Signal, Drought Tolerance and Water Use Efficiency of Winter Wheat[J]. >Journal of Integrative Agriculture, 2013, 12(6): 989-998.

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