疏风解毒胶囊对甲型流感病毒H1N1特异性抗体的影响

doi:10.19803/j.1672-8629.20250170

摘要/Abstract

摘要： 目的探讨疏风解毒胶囊（SFJD）对甲型流感病毒（H1N1）感染小鼠特异性抗体分泌的影响,并通过体外模型研究其对B细胞激活的调控机制。方法建立H1N1非致死性感染小鼠模型,随机分为正常组,PR8模型组,阳性药磷酸奥司他韦组（27.5 mg·kg^-1·d^-1）,SFJD高、中剂量组（2.288 g·kg^-1·d^-1、1.144 g·kg^-1·d^-1）,并将各组小鼠随机分为感染第0、2、4、6、8天共计5个时间点组。第0至第4天进行灌胃给药,于以上时间点对各组小鼠取材,分别称量各组小鼠肺重和体重,计算肺指数;采用高通量液相蛋白芯片多因子法,检测肺泡灌洗液中的总IgA、IgM、IgG;采用酶联免疫吸附法（ELISA）,检测H1N1特异性IgA、IgM与IgG。于第8天,采用实时荧光定量聚合酶链式反应（Real-Time PCR）法,检测各组小鼠肺组织病毒载量;使用Micro-CT,观察小鼠肺部影像;采用流式分析法,检测外周血和肺泡灌洗液B细胞百分比。将人肺上皮细胞（BEAS-2B）分为正常对照组,病毒感染组,阳性药组（62.5μmol·L^-1）,SFJD高、中剂量组（15.6、7.8μg·mL^-1）;小鼠肺上皮细胞（MLE12）分为正常对照组,病毒感染组,阳性药组（250μmol·L^-1）,SFJD高、中剂量组（62.5、31.3μg·mL^-1）。采用蛋白免疫印迹（Western Blot）法,检测SFJD对呼吸道上皮细胞B细胞刺激因子（BAFF）表达的影响。结果 SFJD可降低第4天至第8天H1N1感染小鼠的肺指数（P<0.05）;降低第8天病毒载量（P<0.01）,减轻炎症病变,升高外周血及肺组织中的B细胞比例（P<0.01,P<0.001）;降低第 8天肺泡灌洗液中总IgA水平,升高总IgM水平（P<0.01）;降低第4天肺泡灌洗液中特异性IgA（P<0.000 1）的分泌、促进第8天特异性IgM（P<0.000 1）、IgG（P<0.000 1）的分泌;同时,SFJD可抑制呼吸道上皮细胞系BAFF表达（P<0.05,P<0.001）。结论 SFJD在H1N1感染早期给药,可恢复外周血B细胞比例,促进其向肺组织浸润,增强病毒特异性IgM和IgG分泌,发挥抗病毒效果。同时,SFJD通过抑制呼吸道上皮细胞BAFF表达,防止B细胞过度激活,进而调节感染后的体液免疫应答。

关键词: 甲型流感病毒H1N1, 病毒性肺炎, 疏风解毒胶囊, B细胞刺激因子, 特异性抗体

Abstract: Objective To explore the effects of Shufeng Jiedu capsules (SFJD) on the humoral immune response in mice with non-lethal influenza A virus (H1N1) infection and the mechanisms using an in vitro model. Methods A mouse model of non-lethal H1N1 infection was established. Mice were randomly divided into the normal, PR8 model, positive drug oseltamivir-phosphate (27.5 mg·kg^-1·d^-1), SFJD medium-dose (1.144 g·kg^-1·d^-1), and SFJD high-dose groups (2.288 g·kg^-1·d^-1). Each group was then divided into five time-point subgroups: Days 0, 2, 4, 6, and 8. The drug was administered by gavage from day 0 to day 4. At each time point, the mice were euthanized for sample collection. Lung and body weights were measured to calculate the lung index. Total IgA, IgM, and IgG in bronchoalveolar lavage fluid were detected using a high-throughput liquid phase protein chip multiplex assay. H1N1-specific IgA, IgM, and IgG were measured via enzyme-linked immunosorbent assay (ELISA). On day 8, viral loads in lung tissues were assessed by real-time fluorescence quantitative PCR (Real-time PCR). Lung images were obtained using Micro-CT. The percentage of B cells in peripheral blood and bronchoalveolar lavage fluid was analyzed by flow cytometry. Human bronchi epithelial cells (BEAS-2B) were allocated into 6 groups: normal control, virus infection, positive drug (62.5 μmol·L^-1), SFJD high-dose (15.6 μg·mL^-1) and SFJD medium-dose (7.8 μg·mL^-1). Murine lung epithelial-12 cells (MLE12) were divided into five groups: normal control, virus infection, positive drug (250 μmol·L^-1), SFJD high-dose (62.5 μg·mL^-1) and SFJD medium-dose (31.3 μg·mL^-1). Western blot was employed to evaluate the impact of SFJD on expression levels of B cell activating factor (BAFF) in respiratory epithelial cells. Results SFJD reduced the lung index of H1N1 infected mice from day 4 to day 8 (P<0.05), viral load on day 8 (P<0.01), and alleviated inflammatory lesions. It increased the proportion of B cells in peripheral blood and lung tissues (P<0.01, P<0.001), decreased total IgA levels in bronchoalveolar lavage fluid on day 8, increased total IgM levels (P<0.01), and reduced specific IgA secretion on day 4 (P<0.0001) while promoting specific IgM (P<0.0001) and IgG (P<0.0001) secretion on day 8. SFJD also inhibited BAFF expression in respiratory epithelial cell lines (P<0.05, P<0.001). Conclusion Early administration of SFJD in case of H1N1 infection can restore the peripheral blood B cell proportion, promote their infiltration into lung tissues, enhance specific IgM and IgG secretion, and exert antiviral effects. Additionally, SFJD regulates the humoral immune response post-infection by inhibiting BAFF expression in respiratory epithelial cells and by preventing excessive B cell activation.

Key words: Influenza A Virus H1N1, Viral Pneumonia, Shufeng Jiedu Capsule, B-Cell Activating Factor, Specific Antibody

中图分类号:

李欣颖, 包蕾, 李舒冉, 赵荣华, 孙静, 谢丹, 鲍岩岩, 郭姗姗, 崔晓兰, 耿子涵. 疏风解毒胶囊对甲型流感病毒H1N1特异性抗体的影响[J]. 中国药物警戒, 2025, 22(8): 841-850.

LI Xinying, BAO Lei, LI Shuran, ZHAO Ronghua, SUN Jing, XIE Dan, BAO Yanyan, GUO Shanshan, CUI Xiaolan, GENG Zihan. Effect of Shufeng Jiedu Capsules on the Production of Specific Antibodies against Influenza A H1N1 Virus and the Mechanisms[J]. Chinese Journal of Pharmacovigilance, 2025, 22(8): 841-850.

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