Yu Zeyue, Wang Yudong, Sun Jianhui, Zheng Xiaotong, Hao Liyu, Deng Yurong, Li Jianliang, Li Zongyuan, Shan Zhongchao, Li Weidong, Qiao Yuling, Huo Ruili, Xiong Yibai, Huo Hairu, Li Hui, Lin Longfei, Huang Hanhui, Liu Guimin, Wang Aoao, Li Hongmei, Huang Luqi
National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, No. 16 Nanxiaojie, Dongzhimennei Ave., Beijing 100700, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, No. 16 Nanxiaojie, Dongzhimennei Ave., Beijing 100700, China.
Pharmaceuticals (Basel). 2025 May 27;18(6):802. doi: 10.3390/ph18060802.
: Yiqi Zengmian (YQZM) functions as an immunopotentiator by enhancing both cellular and humoral immunity. However, its pharmacodynamic active constituents, particularly those absorbed into the bloodstream, and mechanism of action remain unclear. This study aimed to investigate the immunopotentiating effects and mechanisms of YQZM in mice immunized with the BBIBP-CorV (Beijing Bio-Institute of Biological Products Coronavirus Vaccine). : Serum pharmacochemistry and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) were employed to identify bioavailable components of YQZM. The mice received the BBIBP-CorV twice on days 1 and 14, while YQZM was orally administered for 28 days. Neutralization assays and ELISA quantified antigen-specific antibodies (abs), flow cytometry (FC) and intracellular cytokine staining (ICS) were used to assess immune cell populations and their cytokines, and an enzyme-linked immunospot assay (ELISpot) quantified memory T and B cells (MBs and MTs). To identify underlying mechanisms, network pharmacology, RNA sequencing (RNA-Seq), molecular docking, Western blotting (WB), and quantitative reverse transcription PCR (RT-qPCR) were performed. : YQZM significantly enhanced antigen-specific antibody titers, immune cell proportions, cytokine levels, and memory lymphocyte functions. UPLC-MS/MS analysis identified 31 bioactive compounds in YQZM. KEGG enrichment analysis based on RNA-Seq and network pharmacology implicated the TLR-JAK-STAT signaling pathway in YQZM's immune-enhancing effects. WB and RT-PCR validated that YQZM upregulated the expression of critical nodes in the TLR-JAK-STAT signaling pathway. Furthermore, molecular docking indicated that YQZM's primary active components exhibited strong binding affinity for critical proteins. : YQZM effectively enhances vaccine-induced innate and adaptive immunity via a multi-component, multi-target mechanism, among which the TLR-JAK-STAT signaling pathway is a validated molecular target.
益气增免（YQZM）通过增强细胞免疫和体液免疫发挥免疫增强剂的作用。然而，其药效学活性成分，特别是那些吸收入血的成分及其作用机制仍不清楚。本研究旨在探讨YQZM对接种BBIBP-CorV（北京生物制品研究所新型冠状病毒疫苗）小鼠的免疫增强作用及其机制。采用血清药物化学和超高效液相色谱-串联质谱（UPLC-MS/MS）法鉴定YQZM的生物可利用成分。小鼠在第1天和第14天接受两次BBIBP-CorV接种，同时YQZM口服给药28天。中和试验和ELISA定量抗原特异性抗体（abs），流式细胞术（FC）和细胞内细胞因子染色（ICS）用于评估免疫细胞群体及其细胞因子，酶联免疫斑点试验（ELISpot）定量记忆T和B细胞（MBs和MTs）。为了确定潜在机制，进行了网络药理学、RNA测序（RNA-Seq）、分子对接、蛋白质免疫印迹（WB）和定量逆转录PCR（RT-qPCR）。YQZM显著提高了抗原特异性抗体滴度、免疫细胞比例、细胞因子水平和记忆淋巴细胞功能。UPLC-MS/MS分析鉴定出YQZM中的31种生物活性化合物。基于RNA-Seq和网络药理学的KEGG富集分析表明TLR-JAK-STAT信号通路与YQZM的免疫增强作用有关。WB和RT-PCR验证YQZM上调了TLR-JAK-STAT信号通路中关键节点的表达。此外，分子对接表明YQZM的主要活性成分对关键蛋白具有很强的结合亲和力。YQZM通过多成分、多靶点机制有效增强疫苗诱导的先天性和适应性免疫，其中TLR-JAK-STAT信号通路是一个经过验证的分子靶点。
