Table 2_Garcinone C inhibits pseudorabies virus replication through EGF/PI3K/Akt axis.xls

IntroductionPseudorabies virus (PRV), a significant pathogen of swine, causes substantial economic losses, and even poses an emergent public health concern due to its zoonotic potential. The continuous evolution of PRV has undermined the effectiveness of current vaccines and antiviral drugs. Consequently, there is an urgent need to develop novel strategies to curb its spread.
MethodsThe inhibitory effect of garcinone C on PRV replication was assessed in vivo and in vitro. To determine the stage of antiviral action, treatments were administered at different time points: pre-treatment, co-treatment, and post-infection. RNA sequencing was performed, and differentially expressed genes (DEGs) were identified.
ResultsIn the study, we found that garcinone C inhibited PRV replication in a concentration- and timedependent manner in vitro. The antiviral activity of garcinone C was specific to post-infection administration and did not extend to pre-treatment and cotreatment conditions. Transcriptomic analysis identified DEGs between garcinone C- and vehicle-treated cells after PRV infection. KEGG pathway enrichment analysis of the DEGs indicated that the antiviral effect of garcinone C was primarily associated with the PI3K-Akt signaling pathway, potentially through the downregulation of the host epidermal growth factor. Furthermore, garcinone C suppressed the production of key inflammatory cytokines such as IL-6, IL-8, and TNF-a during PRV infection. Oral administration of garcinone C conferred protection in PRVinfected mice, leading to attenuated weight loss, an improved survival rate, as well as reduced pathological changes and viral loads in tissues.
DiscussionCollectively, our findings identify garcinone C as a promising therapeutic candidate against PRV and elucidate its underlying molecular mechanism.

引言：伪狂犬病病毒（Pseudorabies virus, PRV）是一类重要的猪源病原体，不仅可造成巨额经济损失，还因其具备人畜共患潜力而成为突发公共卫生隐患。该病毒的持续演化已削弱了现有疫苗与抗病毒药物的防控效果，因此亟需开发全新策略以遏制其传播。
方法：本研究通过体内与体外实验评估了藤黄酮C（garcinone C）对PRV复制的抑制作用。为明确其抗病毒作用阶段，实验设置了不同给药时点：预处理组、共处理组与感染后给药组。本研究开展了RNA测序（RNA sequencing）并筛选得到差异表达基因（differentially expressed genes, DEGs）。
结果：本研究发现，藤黄酮C在体外可通过浓度依赖性与时间依赖性方式抑制PRV复制，且其抗病毒活性仅在感染后给药时显现，预处理与共处理给药均未体现抗病毒效果。转录组分析显示，PRV感染后，藤黄酮C处理组与溶剂对照组细胞的差异表达基因存在显著差异。对差异表达基因进行KEGG通路富集分析后发现，藤黄酮C的抗病毒作用主要与PI3K-Akt信号通路相关，其潜在机制可能为下调宿主表皮生长因子。此外，藤黄酮C可抑制PRV感染过程中IL-6、IL-8及TNF-α等关键炎症细胞因子的产生。藤黄酮C口服给药可对PRV感染小鼠提供保护，具体表现为体重丢失减轻、存活率提升，同时可缓解组织病理损伤并降低组织内病毒载量。
讨论：综上，本研究结果表明藤黄酮C是一种极具潜力的抗PRV候选治疗药物，并阐明了其发挥抗病毒作用的潜在分子机制。