桑树在中国、日本、韩国等亚洲国家广泛用作民族药和功能食品。桑树具有抗病毒、抗氧化、抗胆固醇、抗癌、降血糖、神经保护等多种药理活性。桑树已表现出对流感病毒、SARS-CoV-2等病毒的抗病毒功效，但其抗伪狂犬病病毒（PRV）的潜在活性仍不确定。本研究致力于深入研究桑树的抗伪狂犬病病毒（PRV）潜力。桑叶乙醇提取物（MLE），同时阐明其潜在作用机制。通过qPCR和免疫印迹法评估不同浓度桑树提取物的抗PRV活性。通过 qPCR 和间接免疫荧光测定检测 MLE 在三种不同治疗模式（预处理、联合治疗和后治疗）中对 PRV 复制的抑制作用。 qPCR 用于研究 MLE 对 PRV 感染细胞中 PRV 附着、进入和细胞因子表达的影响。通过UPLC-MS/MS对MLE中的化学成分进行分析。MLE以剂量依赖的方式显着抑制PRV复制和蛋白表达。 MLE 在三种不同的治疗模式下显示出对 PRV 的抑制作用。 MLE在联合治疗模式下的抑制效果最为显着，抑制率为99.42%。 MLE 在早期阶段抑制 PRV 感染。 MLE 通过影响病毒附着和病毒进入来抑制 PRV 感染。此外，MLE 通过减轻 PRV 引发的细胞因子（TNF-α 和 IFN-α）表达升高来抑制 PRV 复制。对其化学成分的分析表明，酚酸和黄酮类化合物是MLE的主要成分。结果表明，MLE通过抑制病毒吸附和进入，同时抑制抗病毒细胞因子的表达，有效阻止PRV感染。因此，MLE 可能是开发治疗人类和动物 PRV 感染的新药物的潜在资源。版权所有 © 2024。由 Elsevier B.V. 出版。

Morus alba L. are widely used as ethnomedicine and functional food in China, Japan, Korea and other Asian countries. Morus alba L. have a variety of pharmacological activity such as antiviral, antioxidation, anti-cholesterol, anticancer, hypoglycemia, and neuroprotection. Morus alba L. has demonstrated antiviral efficacy against influenza viruses, SARS-CoV-2 and so on, but its potential activity against pseudorabies virus (PRV) remains uncertain.This study endeavors to delve into the anti-pseudorabies virus (PRV) potential of the ethanol extract of Morus alba L. leaves (MLE), while simultaneously elucidating its underlying mechanism of action.The anti-PRV activities of Morus alba L. extracts at different concentrations were evaluated by qPCR and immunoblotting. The inhibitory effects of MLE on PRV replication in three distinct treatment modes (pretreatment, co-treatment, and post-treatment) were detected by qPCR and indirect immunofluorescence assays. qPCR was used to investigate the effects of MLE on PRV attachment, entrance, and cytokine expression in PRV-infected cells. The chemical components in MLE were analyzed by UPLC-MS/MS.MLE significantly inhibits PRV replication and protein expression in a dose-dependent manner. MLE displays inhibitory effects against PRV at three different modes of treatment. The most significant inhibitory effect of MLE was observed when used in co-treatment mode, resulting in an inhibition rate of 99.42%. MLE inhibits PRV infection in the early stage. MLE inhibits PRV infection by affecting viral attachment and viral entry. Furthermore, MLE exerts its inhibition on PRV replication by mitigating the heightened expression of cytokines (TNF-α and IFN-α) triggered by PRV. Analysis of its chemical composition highlights phenolic acids and flavonoids as the principal constituents of MLE.The results illustrate that MLE effectively impedes PRV infection by suppressing viral adsorption and entry, while also curbing the expression of antiviral cytokines. Therefore, MLE may be a potential resource for creating new medications to treat human and animal PRV infections.Copyright © 2024. Published by Elsevier B.V.