Luan Hongliang, Song Yizhen, Hu Hongqiao, Zhang Wenrui, Zhang Hui, Su Tianli, Wang Juan, Ye Gang, Yin Zhongqiong, Zhao Xinhong, Zhou Xun, Li Lixia, Zou Yuanfeng, Zhang Yingying, Song Xu
Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China; Qilu Animal Health Products Co., Ltd, Jinan 250100, China.
Natural Medicine Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China.
J Proteomics. 2025 Jun 15;317:105444. doi: 10.1016/j.jprot.2025.105444. Epub 2025 Apr 22.
Pseudorabies virus (PRV) can infect most mammals and has caused significant economic losses in global pig production. The emergence of new mutants significantly reduces the protective effect of vaccination, indicating an urgent need for the development of specific therapeutic agents against PRV infection. In this study, we analyzed the changes in the cellular proteome after PRV infection in resveratrol-treated PK-15 cells using TMT quantitative proteomics combined with LC-MS/MS. The results identified the differential proteins osteopontin (iOPN) and interleukin-1 receptor accessory protein (IL-1RAP), which have significant biological implications. The regulation of OPN-IL-1β signaling by PRV infection was further studied through the OPN-ERK/JNK-IL-1β signaling axis. The transcriptional levels of OPN, C-JUN, IL-1RAP, and IL-1β, along with the protein levels of ERK, JNK, C-Jun, and their phosphorylated forms at 8, 12, and 16 h post-infection, were determined. The results showed that PRV infection inhibited the activation of this signaling axis, which was upregulated by resveratrol treatment. Down-regulation of OPN by siRNA increased PRV proliferation and inhibited the activation of the signaling axis, which was antagonized by resveratrol treatment. In PRV-infected mice, resveratrol treatment produced the same changes observed in vitro. The present study demonstrated that resveratrol can promote innate immune responses by regulating the OPN-ERK/JNK-IL-1β signaling axis, thereby activating host antiviral defenses against PRV infection. SIGNIFICANCE: Resveratrol targets the OPN-ERK/JNK-IL-1β axis to enhance innate immunity, offering a novel antiviral strategy against PRV infection. This study identifies OPN as a key regulator of host defense, linking ERK/JNK signaling to IL-1β-mediated antiviral responses. In vivo validation demonstrates resveratrol's therapeutic potential, reducing PRV replication and mortality in mice via immune pathway activation.
伪狂犬病病毒（PRV）可感染大多数哺乳动物，并在全球养猪业中造成了重大经济损失。新突变体的出现显著降低了疫苗接种的保护效果，这表明迫切需要开发针对PRV感染的特异性治疗药物。在本研究中，我们使用TMT定量蛋白质组学结合LC-MS/MS分析了白藜芦醇处理的PK-15细胞在PRV感染后的细胞蛋白质组变化。结果鉴定出具有重要生物学意义的差异蛋白骨桥蛋白（iOPN）和白细胞介素-1受体辅助蛋白（IL-1RAP）。通过OPN-ERK/JNK-IL-1β信号轴进一步研究了PRV感染对OPN-IL-1β信号的调节。测定了感染后8、12和16小时OPN、C-JUN、IL-1RAP和IL-1β的转录水平，以及ERK、JNK、C-Jun及其磷酸化形式的蛋白水平。结果表明，PRV感染抑制了该信号轴的激活，而白藜芦醇处理可使其上调。用siRNA下调OPN可增加PRV增殖并抑制信号轴的激活，而白藜芦醇处理可拮抗这种作用。在PRV感染的小鼠中，白藜芦醇处理产生了与体外观察到的相同变化。本研究表明，白藜芦醇可通过调节OPN-ERK/JNK-IL-1β信号轴促进先天免疫反应，从而激活宿主针对PRV感染的抗病毒防御。意义：白藜芦醇靶向OPN-ERK/JNK-IL-1β轴以增强先天免疫力，为对抗PRV感染提供了一种新的抗病毒策略。本研究确定OPN是宿主防御的关键调节因子，将ERK/JNK信号与IL-1β介导的抗病毒反应联系起来。体内验证证明了白藜芦醇的治疗潜力，通过激活免疫途径降低小鼠体内PRV的复制和死亡率。
