Yuan Guoqi, Liu Yang, Hong Zhou, Zhu Haiyan, Lu Yan, Chen Daofeng
Department of Natural Medicine, School of Pharmacy, Fudan University, Shanghai, 201203, PR China.
Department of Biological Medicines, School of Pharmacy, Fudan University, Shanghai, 201203, PR China.
J Ethnopharmacol. 2025 Aug 29;352:120222. doi: 10.1016/j.jep.2025.120222. Epub 2025 Jun 28.
Bubali Cornu (BC) has a long history of use in traditional Chinese medicine for the treatment of various conditions, including high fever, hemoptysis, epistaxis, and infections. Despite its historical use, the active components responsible for its therapeutic effects, particularly in viral pneumonia, remain poorly understood.
The objective of this study was to identify the active compounds in BC with antiviral activity against H1N1 and to evaluate their therapeutic potential in alleviating inflammation in H1N1-infected acute lung injury (ALI) mice.
The antiviral components of BC were isolated using silica gel column chromatography and high-performance liquid chromatography (HPLC) according to a bioactive guided separation strategy. Then, the protective effects of these components in H1N1-induced ALI mice were evaluated. To explore the potential mechanisms, network pharmacology (NP) analysis and qPCR were used to screen the related signaling pathways or proteins. Finally, the functions of the most effective compound were verified in H1N1-infected RAW264.7 macrophages, focusing on its regulatory effects on key proteins.
The ethyl acetate fraction (EAF) of BC had a protective effect against ALI induced by H1N1 in mice. Subsequently, 13 compounds were isolated from EAF of BC, of which nicotinic acid (NA) showed the best antiviral activity in vitro. In vivo studies showed that NA treatment of ALI mice infected with H1N1 significantly reduced lung index and inflammation, and improved lung tissue morphology. Flow cytometry analysis revealed that NA reduced the over-recruitment of macrophages and neutrophils to alleviate the inflammatory storm. As verified by NP and qPCR, NA reduced the mRNA levels of NOS2 and NOX1 in the lungs of mice with viral pneumonia, but Western blot analysis only showed that NA inhibited the expression of NOS2. Finally, NA significantly inhibited the expression of NOS2 in RAW264.7 cells infected with H1N1, suggesting that NA might alleviate ALI induced by H1N1 by inhibiting NOS2 expression in pulmonary macrophages.
The present study demonstrated that NA isolated from BC exhibited significant antiviral and anti-inflammatory properties in H1N1-induced ALI. NA might alleviate the progression of H1N1 pneumonia by inhibiting virus replication, reducing the over-recruitment of macrophages and neutrophils, and inhibiting the expression of NOS2. These findings provided valuable insights into the potential of BC in the treatment of viral infections, and highlighted the importance of exploring traditional animal-derived TCM in modern antiviral applications.
水牛角在传统中医中有着悠久的应用历史，可用于治疗多种病症，包括高热、咯血、鼻出血和感染。尽管其有历史应用，但对其治疗作用的活性成分，尤其是在病毒性肺炎方面，仍了解甚少。
本研究的目的是鉴定水牛角中对甲型H1N1流感病毒具有抗病毒活性的活性化合物，并评估它们在减轻甲型H1N1流感病毒感染所致急性肺损伤（ALI）小鼠炎症方面的治疗潜力。
根据生物活性导向分离策略，采用硅胶柱色谱和高效液相色谱（HPLC）分离水牛角的抗病毒成分。然后，评估这些成分对甲型H1N1流感病毒诱导的ALI小鼠的保护作用。为探究潜在机制，运用网络药理学（NP）分析和qPCR筛选相关信号通路或蛋白质。最后，在甲型H1N1流感病毒感染的RAW264.7巨噬细胞中验证最有效化合物的功能，重点关注其对关键蛋白质的调节作用。
水牛角的乙酸乙酯部位（EAF）对甲型H1N1流感病毒诱导的小鼠ALI具有保护作用。随后，从水牛角的EAF中分离出13种化合物，其中烟酸（NA）在体外显示出最佳的抗病毒活性。体内研究表明，用NA治疗甲型H1N1流感病毒感染的ALI小鼠可显著降低肺指数和炎症，并改善肺组织形态。流式细胞术分析显示，NA减少了巨噬细胞和中性粒细胞的过度募集，从而减轻炎症风暴。经NP和qPCR验证，NA降低了病毒性肺炎小鼠肺组织中NOS2和NOX1的mRNA水平，但蛋白质免疫印迹分析仅显示NA抑制了NOS2的表达。最后，NA显著抑制甲型H1N1流感病毒感染的RAW264.7细胞中NOS2的表达，表明NA可能通过抑制肺巨噬细胞中NOS2的表达来减轻甲型H1N1流感病毒诱导的ALI。
本研究表明，从水牛角中分离出的NA在甲型H1N1流感病毒诱导的ALI中具有显著的抗病毒和抗炎特性。NA可能通过抑制病毒复制、减少巨噬细胞和中性粒细胞的过度募集以及抑制NOS2的表达来减轻甲型H1N1流感病毒肺炎的进展。这些发现为水牛角在治疗病毒感染方面的潜力提供了有价值的见解，并强调了在现代抗病毒应用中探索传统动物源中药的重要性。
