Lv Junping, Lin Weishi, Chao Siqi, Xie Jing, Cao Yue, Tie Jinfeng, Ke Yuehua, Lu Binan, Pang Zongran
School of Pharmacy, Minzu University of China, Beijing 100081, China.
Key Laboratory of Ethnomedicine, Minzu University of China, Ministry of Education, Beijing 100081, China.
Pathogens. 2025 Jun 23;14(7):622. doi: 10.3390/pathogens14070622.
Enterovirus A71 (EV-A71), a major etiological agent of hand-foot-mouth disease, can cause severe neurological complications. However, the mechanisms underlying EV-A71-induced cell damage and potential therapeutic strategies remain inadequately understood. Here, we investigated EV-A71 replication dynamics and associated cytopathic effects in nine distinct cell lines, including epithelial, neuronal, immune, and other cell types. Cell viability, membrane integrity, and energy metabolism were assessed using Cell Counting Kit-8 (CCK-8), lactate dehydrogenase (LDH), and adenosine triphosphate (ATP) assays. The antiviral activity of rosmarinic acid (RA), a natural polyphenol, was evaluated by plaque reduction, qPCR, and Western blot. EV-A71 exhibited cell-type-specific replication and cytotoxicity patterns. RA significantly preserved cell viability, reduced LDH release, maintained ATP levels, and suppressed IL-6 expression. Mechanistically, RA inhibited viral replication by downregulating VP1 expression and viral RNA levels. Molecular docking indicated strong binding of RA to the hydrophobic pocket of VP1, potentially disrupting virus-host interactions. Collectively, these findings highlight RA's combined antiviral and cytoprotective potential, supporting its candidacy as a therapeutic agent against EV-A71 infection.
肠道病毒A71（EV - A71）是手足口病的主要病原体，可引发严重的神经并发症。然而，EV - A71诱导细胞损伤的机制以及潜在的治疗策略仍未得到充分了解。在此，我们研究了EV - A71在九种不同细胞系中的复制动态及相关细胞病变效应，这些细胞系包括上皮细胞、神经元细胞、免疫细胞和其他细胞类型。使用细胞计数试剂盒 - 8（CCK - 8）、乳酸脱氢酶（LDH）和三磷酸腺苷（ATP）检测法评估细胞活力、膜完整性和能量代谢。通过噬斑减少试验、定量聚合酶链反应（qPCR）和蛋白质免疫印迹法评估天然多酚迷迭香酸（RA）的抗病毒活性。EV - A71呈现出细胞类型特异性的复制和细胞毒性模式。RA显著维持细胞活力、减少LDH释放、维持ATP水平并抑制白细胞介素 - 6（IL - 6）的表达。从机制上讲，RA通过下调VP1表达和病毒RNA水平来抑制病毒复制。分子对接表明RA与VP1的疏水口袋有强结合，可能破坏病毒与宿主的相互作用。总的来说，这些发现突出了RA的联合抗病毒和细胞保护潜力，支持其作为抗EV - A71感染治疗药物的候选资格。
