Zhang Miao, Huang Jiankun, Chi Qingan, Ran Xuhua, Wen Xiaobo
School of Tropical Agriculture and Forestry, Hainan University, Haikou, Hainan, China.
Hainan Animal Disease Prevention and Control Center, Haikou, Hainan, China.
Front Microbiol. 2025 Apr 29;16:1539094. doi: 10.3389/fmicb.2025.1539094. eCollection 2025.
Currently, vaccination has consistently posed challenges in preventing the Porcine reproductive and respiratory syndrome virus (PRRSV), so there is an urgent need for effective controlling strategies. Ma-Xing-Shi-Gan-San (MXSGS), a traditional Chinese medicine (TCM) formula used for pulmonary diseases and respiratory disorders, has proven effective in treating H1N1 and COVID-19. Herein, we evaluated whether MXSGS exhibits potent antiviral activity against PRRSV.
First, a PRRSV-infected Marc-145 cell model was established. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and the tissue culture infective dose (TCID₅₀) assay were performed to assess the inhibitory effects of MXSGS on PRRSV during different administration stages. Network pharmacology was then employed to identify key active ingredients and core potential targets of MXSGS against PRRSV. In addition, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to elucidate the antiviral signaling pathways modulated by MXSGS. Lastly, candidate ingredients and targets were validated by molecular docking analysis.
MXSGS significantly inhibited PRRSV through prophylactic and therapeutic administration and suppressed multiple phases of the viral life cycle, including attachment, internalization, replication, and release. In network pharmacology results, 82 active ingredients and 118 therapeutic targets related to MXSGS and PRRSV were identified. Among them, Calycosin, Odoratin, Glyzaglabrin, 7,2',4'-trihydroxy-5-methoxy-3-arylcoumarin, and Eriodictyol were selected as key active ingredients. ALB, PPARG, CASP3, STAT3, TGFB1, JAK2, TLR4, PRKACA, and PRKACB were screened as potential core targets. Furthermore, pathway and functional enrichment analysis revealed that the impact of MXSGS on PRRSV mainly involved Toll-like receptor signaling pathway, typical NF-κB signaling, positive regulation of interleukin-6 production, Th17 cell differentiation, inflammatory response, and viral defense response. Lastly, molecular docking analysis indicated an excellent binding affinity between the core potential targets and key active ingredients, with all binding energies < -6.0 kcal/mol.
experiments indicated that MXSGS exhibited considerable anti-PRRSV activity. Using network pharmacology and molecular docking approaches, five key active ingredients and six core potential targets were identified, underscoring MXSGS as a promising pharmaceutical agent for controlling PRRSV.
目前，疫苗接种在预防猪繁殖与呼吸综合征病毒（PRRSV）方面一直面临挑战，因此迫切需要有效的控制策略。麻杏石甘散（MXSGS）是一种用于治疗肺部疾病和呼吸紊乱的传统中药配方，已被证明对治疗H1N1和COVID - 19有效。在此，我们评估了MXSGS是否对PRRSV具有强大的抗病毒活性。
首先，建立PRRSV感染的Marc - 145细胞模型。进行逆转录定量聚合酶链反应（RT - qPCR）和组织培养感染剂量（TCID₅₀）测定，以评估MXSGS在不同给药阶段对PRRSV的抑制作用。然后采用网络药理学方法确定MXSGS抗PRRSV的关键活性成分和核心潜在靶点。此外，进行基因本体（GO）和京都基因与基因组百科全书（KEGG）分析，以阐明MXSGS调节的抗病毒信号通路。最后，通过分子对接分析验证候选成分和靶点。
MXSGS通过预防性和治疗性给药显著抑制PRRSV，并抑制病毒生命周期的多个阶段，包括附着、内化、复制和释放。在网络药理学结果中，鉴定出82种与MXSGS和PRRSV相关的活性成分和118个治疗靶点。其中，毛蕊异黄酮、欧前胡素、甘草香豆素、7,2',4'-三羟基-5-甲氧基-3-芳基香豆素和圣草酚被选为关键活性成分。筛选出ALB、PPARG、CASP3、STAT3、TGFB1、JAK2、TLR4、PRKACA和PRKACB作为潜在的核心靶点。此外，通路和功能富集分析表明，MXSGS对PRRSV的影响主要涉及Toll样受体信号通路、典型的NF-κB信号通路、白细胞介素-6产生的正调控、Th17细胞分化、炎症反应和病毒防御反应。最后，分子对接分析表明核心潜在靶点与关键活性成分之间具有良好的结合亲和力，所有结合能均 < -6.0 kcal/mol。
实验表明MXSGS具有显著的抗PRRSV活性。通过网络药理学和分子对接方法，确定了五种关键活性成分和六个核心潜在靶点，突出了MXSGS作为控制PRRSV的有前景的药剂。
